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"Diabetes melitus tipe 2 (DMT2) adalah penyakit yang ditandai dengan hiperglikemia dan dikaitkan dengan keadaan inflamasi kronis, Sel punca merupakan terapi sel yang menjanjikan untuk DMT2 namun, efek parakrin dari hasil sekresi sel punca atau sekretom lebih potensial. Sekretom dan Autologous Activated Platelet Rich Plasma (aaPRP) berpotensi sebagai terapi bebas sel dan memiliki kandungan biomolekul yang sama untuk terapi DMT2. Penelitian ini bertujuan untuk mengevaluasi sitokin dan faktor pertumbuhan pada aaPRP dan sekretom sebagai strategi awal pengembangan aaPRP dan sekretom untuk DMT2. Penelitian ini adalah eksperimental in-vitro dengan total 13 pasien (10 DMT2 dan 3 Non DMT2). Analisis total protein dan Luminex terhadap sitokin TNF-α, IL-6, IL-10, growth factor VEGF, dan FGF dilakukan pada plasma, aaPRP, dan sekretom. Sekretom diisolasi dari sel punca jaringan adiposa (ADSC) dengan suplemtasi media kultur PRP Kontrol dan FBS. Hasil penelitian menunjukkan, sitokin dan growth factor dari aaPRP dan sekretom Non DMT2, mayoritas lebih tinggi dibandingkan DMT2. Terdapat perbedaan sitokin dan growth factor antara sekretom dengan media PRP Kontrol dan FBS. Kondisi DMT2 mempengaruhi sitokin dan growth factor pada aaPRP. Sitokin dan growth factor pada sekretom dipengaruhi oleh kondisi DMT2 dan media kultur.

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Type 2 diabetes mellitus (T2DM) is a disease characterized by hyperglycemia and is associated with a chronic inflammatory state. Stem cells are a promising cell therapy for T2DM however, the paracrine effect of stem cell secretions, or the secretome, has more potential. Secretome and Autologous Activated Platelet Rich Plasma (aaPRP) can be cell-free therapies for T2DM. This study aims to evaluate cytokines and growth factors in aaPRP and the secretome as an initial strategy for developing aaPRP and the secretome for T2DM. This was an in-vitro experimental study with 13 patients (10 T2DM and 3 non-T2DM). Total protein and Luminex assays of the cytokines TNF-α, IL-6, IL-10, growth factor VEGF, and FGF were taken from plasma, aaPRP, and the secretome. The secretome was isolated from adipose tissue stem cells (ADSC) by supplementing control PRP and FBS culture media. The results showed that most cytokines and growth factors from aaPRP and the non-DMT2 secretome were higher than those from DMT2. There are differences in cytokines and growth factors between the secretome and control PRP and FBS media. T2DM conditions affect cytokines and growth factors in aaPRP. Cytokines and growth factors in the secretome are influenced by T2DM conditions and culture mediums."

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Latar Belakang: Diabetes melitus (DM) tipe 2 adalah suatu penyakit metabolik yang kompleks dan kronis yang ditandai dengan gangguan angiogenesis. Inflamasi kronis derajat ringan dan stres oksidatif yang meningkat pada DM tipe 2 diketahui dapat menyebabkan gangguan fungsi biologis pada sel progenitor/sel punca vaskular, salah satunya adalah adipose-derived mesenchymal stem cell (ADSC). Sejumlah penelitian menunjukkan potensi vaskulogenik ADSC dan perannya pada regenerasi jaringan. Hingga saat ini aplikasi sel punca autologus pada penderita DM untuk menginisiasi vaskularisasi masih mengalami kendala. Platelet-rich plasma (PRP) diketahui kaya akan berbagai faktor pertumbuhan, termasuk VEGF, yang penting untuk proses angiogenesis.

Tujuan: Penelitian ini bertujuan untuk menguji dan menganalisis efek pemberian PRP PMI terhadap proliferasi (jumlah sel stromal, nilai population doubling time (PDT), dan persentase sel hidup), diferensiasi (pembentukan koloni, ekspresi CD73, CD90, CD105, dan tiga lini diferensiasi), ekspresi mRNA VEGF dan VEGFR2, dan potensi angiogenik ADSC DM in vitro (sekresi VEGF dan pembentukan tubular kapiler).

Metode: Terlebih dahulu, konsentrasi trombosit per µL dan kadar VEGF per 1x10³ trombosit pecah yang terkandung dalam PRP Palang Merah Indonesia (PMI) dibandingkan dengan PRP DM dan non-DM. Lalu, stromal vascular fraction (SVF) diisolasi dari jaringan lemak menggunakan metode enzimatik, dan SVF penderita DM tipe 2 (n= 15) dan non-DM (n= 10)  dikultur dalam medium kontrol hingga didapat ADSC pasase 1−3 (P1−P3). Proliferasi, diferensiasi, ekspresi mRNA VEGF dan VEGFR2, serta potensi angiogenik ADSC DM dan non-DM diukur dan dibandingkan. ADSC DM P3 kemudian dikultur dalam medium PRP PMI 5%, 10%, 15%, dan 20%, lalu proliferasi, diferensiasi, ekspresi mRNA VEGF dan VEGFR2 diukur dan dibandingkan dengan kontrol (FBS) untuk mendapatkan konsentrasi PRP optimum. ADSC DM P3 yang diprekondisikan dengan PRP optimum, dengan atau tanpa anti-VEGF (bevacizumab) 100 ng/mL, dan ADSC DM P3 kontrol dikultur, lalu sekresi VEGF dan pembentukan tubular kapiler pada Matrigel^® diukur.

Hasil: Pada penelitian ini tidak ditemukan perbedaan bermakna antara konsentrasi trombosit per µL PRP DM, non-DM, dan PMI (p= 0,22). Namun, PRP non-DM memiliki kadar VEGF per 1000 trombosit pecah lebih rendah bermakna (0,20 (0,04−0,35) fg) dibandingkan PRP DM (0,69 (0,21−1,17) fg), p= 0,03) dan PMI (1,84 (1,38−2,10) p= 0,01), dan tidak ada perbedaan bermakna antara PRP DM dan PRP PMI (p= 0,06). Jumlah sel stromal per gram lemak dan jumlah koloni sel stromal DM lebih rendah dari non-DM (86,35 (52,48−106,76) x 10⁶ vs 158,93 (101,59−185,94) x 10⁶, p= 0,01, dan 94 ± 14 koloni vs 31 ± 32 koloni, p= 0,004). Tidak terdapat perbedaan bermakna antara DM dan non-DM pada persentase sel stromal hidup (p= 0,24), ekspresi CD73 (p= 0,21), CD90 (p= 0,90), adipogenesis, kondrogenesis, osteogenesis, PDT P2 (p= 0,27), PDT P3 (p= 0,21), dan persentase sel hidup ADSC P2 (p= 0,07), sedangkan ekspresi CD105 (64,41 (51,20−73,38)% vs 91,40 (82,62−95,47)%, p< 0,001) ADSC DM P1 dan persentase sel hidup (82,70 ± 8,07% vs 91,15 ± 3,77%, p= 0,04) ADSC DM P3 lebih rendah bermakna dibandingkan ADSC non-DM. Tidak ada penurunan yang bermakna pada ekspresi relatif mRNA VEGF (0,64 (0,30−1,08), p= 0,86) dan VEGFR2 DM (0,64 ± 0,56, p= 0,49) jika dibandingkan dengan ADSC non-DM. Rerata kadar VEGF dalam conditioned medium (CM) yang disekresikan oleh 1x10³ ADSC DM dan non-DM secara berturut-turut sebesar 0,74 pg/mL dan 0,62 pg/mL. ADSC DM yang diberi PRP optimum, yaitu 15% memiliki nilai PDT yang lebih rendah (2,33 ± 0,56 hari vs 5,04 ± 1,26 hari, p= 0,01) dan persentase sel hidup (95,53 ± 1,60% vs 78,95 ± 10,13%, p=0,01) yang lebih tinggi bermakna dibandingkan dengan kontrol. Terjadi peningkatan ekspresi CD105, mRNA VEGF, dan VEGFR2 ADSC DM yang diberi PRP 15% (secara berturut-turut 1,81 ± 0,73, p= 0,01; 5,27 ± 5,69, p= 0,23; dan 9,01 ± 11,59, p= 0,06) relatif terhadap kontrol. ADSC DM yang diberi PRP 15% dan ADSC DM kontrol secara berturut-turut mensekresikan VEGF rerata sebanyak 0,57 pg/mL dan 1,67 pg/mL per 1x10³ sel hidup. Jumlah tubular kapiler in vitro ADSC DM yang diberi PRP meningkat pada jam ke-24 jika dibandingkan dengan kontrol dan tidak berbeda bermakna dengan ADSC non-DM, namun membutuhkan waktu lebih panjang, serta tidak berbeda bermakna dengan ADSC DM yang diberi PRP dan anti-VEGF (p=0,78).

Kesimpulan: ADSC DM terbukti mengalami kerusakan selular yang dicirikan dengan penurunan proliferasi, diferensiasi, ekspresi mRNA VEGF dan VEGFR2, serta potensi angiogeniknya. Pemberian PRP 15% (VEGF 98,00 pg/mL) dapat memperbaiki kerusakan tersebut melalui efek sinergis yang dihasilkan oleh VEGF dan faktor pertumbuhan lainnya yang terdapat dalam PRP.

 

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Background: Type II diabetes mellitus (DM type 2) is a chronic and complex metabolic disease identified by impaired angiogenesis. Low grade chronic inflammation and increasing oxidative stress in DM type 2 decrease the biological functions of progenitor/stem cells, including adipose-derived stem cells (ADSC).  ADSC plays significant roles in angiogenesis and tissue regeneration. Some studies have shown the vasculogenic potency of ADSC and its role in tissue regeneration. To date, autologous cell application in DM patients to initiate vascularization is hindered. Platelet-rich plasma (PRP) is widely known to contain generous amount of growth factors including VEGF with significant role in angiogenesis.

Objective: This study aimed to investigate and analyze the effect of PRP preconditioning to the proliferation (stromal cell number, population doubling time (PDT) and percentage of viable cells), differentiation (colony formation, CD73, CD90, CD105 expressions, three lineage of differentiation), expression of mRNA VEGF and VEGFR2, as well as in vitro angiogenic potency of ADSC DM (VEGF secretion and capillary tube formation).

Methods: Initially, platelet concentration per µL and VEGF per 1x10³ lysed platelet contained in Palang Merah Indonesia (PMI) PRP was compared to DM and non-DM PRP. Subsequently, stromal vascular fraction (SVF) from 15 DM and 10 non-DM donors was enzymatically isolated from adipose tissue, and cultured in control media to generate passage 1−3 (P1−P3) ADSC. Proliferation, differentiation, mRNA VEGF and VEGFR2 expression, as well as angiogenic potency of DM ADSC in vitro were measured and compared to non-DM control. P3 DM ADSC was then cultured in media contained 5%, 10%, 15%, and 20% PMI PRP, and proliferation, differentiation, mRNA VEGF and VEGFR2 expression were measured and compared to FBS control to determine optimum PMI PRP concentration. P3 DM ADSC preconditioned with optimum PMI PRP, with or without anti-VEGF (bevacizumab) 100 ng/mL, and control DM ADSC were cultured, and VEGF secretion was measured, as well as capillary tube formation on Matrigel^®.

Results: In this study no significant differences were observed between platelet concentration per µL DM, non-DM, and PMI PRP (p= 0.22). However, non-DM PRP contained significantly lower VEGF per 1000 lysed platelets (0.20 (0.04−0.35) fg) compared to DM (0.69 (0.21−1.17) fg, p= 0.03) and PMI PRP (1.84 (1.38−2.10), p= 0.01), with no significant difference between DM and PMI PRP (p=0.06). The number of viable stromal cells per gram adipose tissue and collonies generated from DM SVF were significantly lower than non-DM (86.35 (52.48−106.76) x 10⁶ vs 158.93 (101.59−185.94) x 10⁶, p= 0.01 and 94 ± 14 collonies vs 31 ± 32 collonies, p= 0.004). Non-significant differences were also observed in the percentage of viable stromal cells (p= 0.24), expression of CD73 (p= 0.21), CD90 (p= 0.90), adipogenesis, chondrogenesis, osteogenesis, P2 and P3 PDT (p= 0.27 and 0.21, respectively), and the percentage of viable P2 ADSC (p= 0.07), but the expression of CD105 of P1 DM ADSC (64.41 (51.20−73.38)% vs 91.40 (82.62−95.47)%, p< 0.001) and the percentage of viable P3 ADSC (82.70 ± 8.07% vs 91.15 ± 3.77%, p= 0.04) were significantly lower than non-DM ADSC. The reduction of mRNA VEGF and VEGFR2 relative expression of P3 DM ADSC (0.64 (0.30−1.08) p= 0.86 and 0.64 ± 0.56, p= 0.49, respectively) were unsignificant compared to non-DM ADSC. Mean of VEGF level normalized to 1x10³ viable cells in the conditioned medium (CM) of DM and non-DM ADSC were 0.74 pg/mL and 0.62 pg/mL, respectively. Optimum 15% PRP-preconditioned DM ADSC had significantly lower PDT value (2.33 ± 0.56 days vs 5.04 ± 1.26 days, p=0.01) and higher percentage of viable cells compared to control (95.53 ± 1.60% vs 78.95 ± 10.13%, p=0.01). The increase of relative expression of CD105, mRNA VEGF and VEGFR2 (1.81 ± 0.73, p= 0.01; 5.27 ± 5.69, p= 0.23; and 9.01 ± 11.59, p= 0.06, respectively) were unsignificant in DM ADSC compared to non-DM. Optimum 15% PRP-preconditioned DM ADSC and control secreted VEGF in CM as much as 0.57 pg/mL and 1.67 pg/mL per 1x10³ viable cells in average. PRP-preconditioning improved the capillary tube formation in DM ADSC, but the process was longer compared to control, and unsignificant to non-DM ADSC and PRP-preconditioned DM ADSC with anti-VEGF (p=0.78).

Conclusions: Cellular damage in DM ADSC was idientified by a reduction of proliferation, differentiation. mRNA VEGF and VEGFR2 expression, and angiogenic potency. Preconditioning DM ADSC with 15% PRP (VEGF 98.00 pg/mL) improved the cellular damage with synergitic effect of VEGF and other growth factors contained in PRP.

 

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"Latar Belakang: pulpa memiliki sifat low-compliance yang memengaruhi proses regenerasinya. Tujuan: menganalisis potensi Platelet- Rich Plasma (PRP) Eksosom terhadap regenerasi pulpa gigi secara in-vitro viabilitas sel, aktivitas migrasi, dan ekspresi Vascular Endothelial Growth Factor-A (VEGF-A) hDPSCs. Metodologi: hDPSC sembilan gigi molar tiga dikultur dengan metode enzyme digestion (ED) yang dipanen pada P3 dan P4. Kemudian dikultur di dalam enam media, yaitu: Dulbecco's Modified Eagle Medium (DMEM) dan 10% PRP sebagai kelompok kontrol, dan 0,5%, 1%, dan 5% eksosom dari PRP. Semua kelompok memiliki tiga rangkap biologis (Triplo). Uji viabilitas sel dievaluasi dengan MTT assay, aktivitas migrasi sel dengan Scratch Assay dan Transwell Migration Assay, dan ekspresi VEGF-A dengan Enzyme-Linked Lmmunosorbent Assay (ELISA). Analisis data dilakukan dengan uji One Way ANOVA (p <0,05) serta uji Kruskal-Wallis dan post hoc Mann-Whitney (p <0,05). Hasil: viabilitas hDPSCs tertinggi pada 24, 48 dan 72 jam observasi pada kelompok Eksosom dari PRP 5% (p <0,05). Eksosom dari PRP 5% menunjukkan aktivitas migrasi yang lebih tinggi dibandingkan dengan kelompok lain, meskipun terdapat perbedaan tidak bermakna dengan kontrol PRP 10% (p >0,05). Ekspresi VEGF-A hDPSCs tertinggi terdapat pada kelompok PRP Eksosom 5% pada 72 jam observasi. Kesimpulan: eksosom dari PRP 5% berpotensi menginduksi regenerasi pupa gigi manusia.

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Background: pulp has low-compliance properties that affect its regeneration process. Objective: to analyze the potential of Platelet-Rich Plasma (PRP) exosomes on the regeneration of dental pulp by in-vitro evaluation of cell viability, migration activity, and expression of Vascular Endothelial Growth Factor-A (VEGF-A) hDPSCs. Methodology: hDPSCs of nine third molars cultured by enzyme digestion (ED) method were harvested at P3 and P4. Then cultured in six media, Dulbecco's Modified Eagle Medium (DMEM) and 10% PRP as a control group, and 0.5%, 1%, and 5% exosomes of PRP. All groups had a biological triple (Triplo). Cell viability assay was evaluated by MTT assay, cell migration activity by Scratch Assay and Transwell Migration Assay, and VEGF-A expression by Enzyme-Linked Lmmunosorbent Assay (ELISA). Data analysis was performed using One Way ANOVA (p < 0.05) and Kruskal-Wallis and post hoc Mann-Whitney tests (p < 0.05). Results: The viability of hDPSCs was highest at 24, 48 and 72 hours of observation in the Exosomes group of 5% PRP (p < 0.05). Exosomes from 5% PRP showed higher migratory activity compared to other groups, although there was no significant difference with 10% PRP control (p > 0.05). The highest expression of VEGF-A hDPSCs was found in the 5% PRP Exosomes group at 72 hours of observation. Conclusion: exosomes of 5% PRP have the potential to induce the regeneration of human dental pulp."

"Pendahuluan: Sekretom sel punca mesenkimal dipercaya mengandung faktor pertumbuhan yang bekerja melalui mekanisme parakrin di situs cedera. Di antara banyaknya faktor-faktor pertumbuhan, beberapa disinyalir memiliki efek osteogenik antara lain bone morphogenetic protein-2 (BMP-2), epidermal growth factor (EGF), dan vascular endothelial growth factor (VEGF).Tujuan: Tujuan penelitian ini adalah untuk mengidentifikasi kuantitas BMP-2, EGF dan VEGF pada sekretom sel punca mesenkimal jaringan adiposa dan tali pusat.Metode: Sampel sekretom dari sel punca mesenkimal jaringan adiposa dan tali pusat dibedakan berdasarkan perlakuan pemberian serum atau non serum dan waktu pengambilan saat penggantian medium terakhir atau saat panen, dan dianalisa dengan metode ELISA sandwich assay menggunakan Human BMP-2, VEGF, and EGF ELISA Kits.Hasil: Sebaran nilai BMP-2 tersentrasi pada nilai 0 pada sekretom jaringan adiposa maupun tali pusat. Kadar EGF dan VEGF memiliki perbedaan bermakna pada sampel jaringan adiposa yang berbeda (p<0,009 dan p<0,005). Kadar EGF dan VEGF pada jaringan adiposa adalah 2,67 (0-22,53) dan 1473,5 (136,1-5335) sedangkan pada jaringan tali pusat adalah 2,67 (0-13,29) dan 0 (0-1675).Kesimpulan: Sekretom jaringan adiposa dan tali pusat kemungkinan hanya mengandung BMP-2 dalam nilai yang sangat rendah. Baik jaringan adiposa maupun jaringan tali pusat mengandung EGF dalam jumlah yang moderat. Kadar VEGF pada jaringan adiposa secara signifikan lebih tinggi.

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Background: The secretome derived from mesenchymal stem cells has been suggested contain growth factors that works via a paracrine mechanism in the injured area. Of these factors, some are thought to have an osteogenic effect, including bone morphogenetic protein-2 (BMP-2), epidermal growth factor (EGF), and vascular endothelial growth factor (VEGF).

Objective: The aim of this study is to identify the quantity of BMP-2, EGF, VEGF in secretome from adipose tissue (AT-MSC) and umbilical cord-derived mesenchymal stem cells (UC-MSC).

Methods: Secretome samples from AT-MSC and UC-MSC were grouped based on serum administration and harvesting time, and were analyzed with an ELISA sandwich assay method using Human BMP-2, VEGF, and EGF ELISA Kits. This study aims to identify whether BMP-2 is contained in the secretome of AT-MSC and UC-MSC, which has never been reported before, and to measure the level of EGF and VEGF within the secretome.

Results: The distribution of value for BMP-2 was nearly zero in the secretome of AT-MSC and UC-MSC. The level of EGF and VEGF were significantly different between different donor samples of AT-MSC (p<0,009 and p<0,005). The level of EGF and VEGF of AT-MSC are 2,67 (0-22,53) and 1473,5 (136,1-5335) compare to 2,67 (0-13,29) and 0 (0-1675) of UC-MSC.

Conclusion: The secretome of AT-MSC and UC-MSC may contain BMP-2 in a very low level. Both AT-MSC and UC-MSC contain EGF in moderate amount. VEGF is significantly higher in of AT-MSC."

"Latar Belakang: Individu dengan diabetes melitus tipe 2 (DMT2) dilaporkan mengalami peningkatan risiko terjadinya sarkopenia dan juga sebaliknya. Penelitian mengenai DMT2 dengan sarkopenia mayoritas berasal dari populasi geriatri. Sampai saat ini belum ada studi yang membandingkan profil metabolik dan parameter inflamasi di kelompok DMT2 dengan dan tanpa sarkopenia pada usia yang lebih muda.Tujuan: Penelitian ini bertujuan untuk mengetahui adanya perbedaan rerata profil metabolik dan parameter inflamasi pada penyandang DMT2 nongeriatri dengan dan tanpa sarkopenia.Metode: Penelitian potong lintang ini melibatkan individu dengan DMT2 nongeriatri berusia  18-59 tahun yang berobat di Rumah Sakit Cipto Mangunkusumo (RSCM), Jakarta, Indonesia pada bulan Januari 2021- Januari 2022. Dilakukan pengambilan data sekunder berupa antropometri dan laboratorium yang mencakup Homeostatic Model Assessment for Insulin Resistance (HOMA-IR), HbA1c dan profil lipid. Kadar interleukin (IL)-6 dan IL-10 serum diukur menggunakan teknik ELISA. Kelompok sarkopenia terdiri atas possible dan true sarcopenia berdasarkan kriteria Asian Working Group for Sarcopenia (AWGS) 2019.Hasil: Dari 100 subjek, 35 subjek dikategorikan ke dalam possible sarkopenia dan 4 subjek true sarkopenia. Subjek DMT2 nongeriatri dengan sarkopenia memiliki median (RIK) nilai HOMA-IR dan kadar HbA1c yang lebih tinggi dibanding subjek tanpa sarkopenia yaitu berturut-turut [6,52 (4,05-17,26) vs. 4,66 (2,61-10,14); p=0,025] dan [9,0% (7,3-10,3)% vs. 7,4% (6,6-8,45)%; p=0,002]. Tidak terdapat perbedaan kadar profil metabolik lain dan IL-6 antara kedua kelompok, sementara kadar IL-10 hanya terdeteksi pada 33 sampel sehingga tidak dapat dianalisis lebih lanjut.Kesimpulan: Median nilai HOMA-IR dan kadar HbA1c kelompok DMT2 nongeriatri dengan sarkopenia lebih tinggi dibanding kelompok tanpa sarkopenia. Tidak ditemukan perbedaan kadar profil metabolik lain dan IL-6 sebagai parameter inflamasi antara kedua kelompok tersebut. Tidak dilakukan analisis beda rerata kadar IL-10 karena sedikitnya sampel yang terdeteksi.

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Background: Individuals with type 2 diabetes mellitus (T2DM) are at increased risk for sarcopenia and vice versa. Studies in T2DM with sarcopenia mostly came from the geriatric population. To date, no study has compared the metabolic profile and inflammatory parameters in younger T2DM subjects with and without sarcopenia.

Aim: This study aimed to assess the mean differences in the metabolic profile and inflammatory parameters of nongeriatric T2DM individuals with vs. without sarcopenia.

Method: This cross-sectional study involved nongeriatric T2DM individuals aged 18-59 years old visiting Cipto Mangunkusumo Hospital, Jakarta, Indonesia between January 2021 and January 2022. Secondary data was obtained, namely anthropometric and laboratory data including Homeostatic Model Assessment for Insulin Resistance (HOMA-IR), HbA1c and lipid profile. Serum levels of interleukin (IL)-6 and IL-10 were measured using the ELISA technique. The sarcopenia group consists of individuals with possible and true sarcopenia based on the Asian Working Group for Sarcopenia (AWGS) 2019 criteria.

Results: From 100 subjects, 35 was categorized as possible sarcopenia and 4 as true sarcopenia. Nongeriatric T2DM subjects with sarcopenia had significantly higher median (interquartile range) HOMA-IR and HbA1c compared to nonsarcopenic subjects [6.52 (4.05-17.26) vs. 4.66 (2.61-10.14); p=0.025] and [9.0% (7.3-10.3)% vs. 7.4% (6.6-8.45)%; p=0.002]. There were no differences in other levels of metabolic profile and IL-6 between the two groups, while IL-10 levels were only detected in 33 samples and could not be analyzed further.

Conclusion: Median HOMA-IR and HbA1c nongeriatric T2DM subjects with sarcopenia was higher than those without sarcopenia. There was no difference in other metabolic profile and IL-6 level as inflammation parameter between the two groups. IL-10 was not analysed further due to the small sample number that were detected."

"Tujuan dari penelitian ini adalah untuk menganalisis pengaruh Platelet-Rich Plasma (PRP) Eksosom terhadap potensi regenerasi jaringan pulpa gigi dengan evaluasiin vitro viabilitas sel, aktivitas migrasi, dan ekspresi Vascular Endothelial Growth Factor-A (VEGF-A) sel punca pulpa gigi manusia (hDPSCs). HDPSC diambil darisembilan gigi molar tiga dari sembilan donor sesuai kriterian inklusi, dan isolasi serta kultur dilakukan dengan metode enzyme digestion (ED) yang dipanen antara P3 dan P4. Setelah starvatation, hDPSCs dikultur di dalam enam media, yaitu sebagai berikut: Dulbecco's Modified Eagle Medium (DMEM) dan 10% PRP sebagai kelompok kontrol, dan 0,5%, 1%, dan 5% PRP eksosom sebagai kelompok eksperimental. Semua kelompok memiliki tiga rangkap biologis (Triplo). Uji viabilitas sel dievaluasi dengan MTT assay, aktivitas migrasi sel dengan Scratch Assay dan Transwell Migration Assay, dan ekspresi VEGF-A dengan Enzyme- Linked Lmmunosorbent Assay (ELISA). Analisis data dilakukan dengan uji One Way ANOVA (p <0,05) serta uji Kruskal-Wallis dan post hoc Mann-Whitney (p<0,05). Nilai rata-rata viabilitas hDPSCs tertinggi pada 24, 48 dan 72 jam observasi pada kelompok PRP-Eksosom 5% (p <0,05). PRP Eksosom 5% menunjukkan aktivitas migrasi yang lebih tinggi dibandingkan dengan kelompok lain, meskipun tidak terdapat perbedaan bermakna dengan kontrol PRP 10% (p> 0,05). Ekspresi VEGF-A hDPSCs tertinggi terdapat pada kelompok PRP Eksosom 5% pada 72 jampengamatan. Dapat disimpulkan bahwa eksosom PRP 5% berpotensi menginduksi regenerasi pupa gigi manusia.

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The purpose of this study was to analyze the effect of Exosome Platelet-Rich Plasma (PRP) on their potential for human Dental Pulp regeneration by evaluating in vitro cell viability, migration activity, and expression of Vascular Endothelial Growth Factor-A (VEGF-A) of human Dental Pulp Stem Cells (hDPSCs). hDPSCs was taken from nine third molars from nine donors thatfit to the inclusion criteria of this study, isolation and culture were carried out by the enzyme digestion (EZ) method harvested between P3 and P4. After starvatation,hDPSCs were cultured in six media, namely as follows: Dulbecco's Modified EagleMedium (DMEM) and 10% PRP as the control group, and 0.5%, 1%, and 5% PRP exosomes as experimental groups. All groups had a biological triplication (Triplo). Cell viability test was evaluated by MTT assay, cell migration activity with Scratch Assay and Transwell Migration Assay, and VEGF-A expression by Enzyme- Linked Immunosorbent Assay (ELISA). Data analysis was performed using One Way ANOVA (p <0.05) and Kruskal-Wallis (p <0.05) and Pearson/Spearman Correlation test (p<0.05). The highest mean hDPSCs viability was at 24, 48 and 72 hours of observation in the PRP-exosome group 5% (p <0.05). Exosome PRP 5% showed higher migration activity compared to other groups, although there was no significant difference with PRP control 10% (p> 0.05). The highest expression of VEGF-A hDPSCs was found in the PRP exosome group 5% at 72 hours of observation. It can be concluded that the PRP 5% exosome have the highest potential ability in inducing pulp regeneration."

"Hiperglikemia kronis yang jika tidak terkontrol dapat merusak pembuluh darah yang merupakan salah satu penyebab utama kematian terkait diabetes yang disebabkan oleh stroke. Penelitian ini bertujuan untuk mengetahui gambaran kesadaran diri penyandang diabetes mellitus terhadap risiko komplikasi stroke di kota depok. Penelitian deskriptif yang melibatkan 100 responden penyandang diabetes melitus tipe 2 di Kota Depok. Pengambilan data secara offline di 3 puskesmas Kota Depok menggunakan kuesioner kesadaran diri terhadap risiko komplikasi stroke yang dikembangkan sendiri dengan nilai alpha cronbach 0,826. Hasil penelitian ini menunjukkan mayoritas berada pada rentang usia 61-80 tahun, berjenis kelamin perempuan serta mayoritas responden menganut agama Islam. Mayoritas responden menempuh pendidikan terakhir tingkat SMA, tidak pernah mengikuti penyuluhan/edukasi DM dan telah menyandang DM selama <5 tahun. Mayoritas responden memiliki kesadaran diri terhadap risiko komplikasi stroke yang sedang. Responden memiliki tingkat kesadaran diri rendah, lebih banyak daripada responden yang telah memiliki kesadaran diri tinggi terhadap risiko komplikasi stroke. Penelitian ini merekomendasikan pemberian edukasi kesehatan terkait risiko komplikasi stroke pada penyandang diabetes melitus tipe 2 agar dapat meningkatkan kesadaran diri terhadap risiko komplikasi stroke.

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Chronic hyperglycemia which if not controlled can damage blood vessels which is one of the leading causes of diabetesrelated death caused by stroke. This study aims to find out the picture of self-awareness of people with diabetes mellitus to the risk of stroke complications in the city of Depok. Descriptive research involving 100 respondents with type 2 diabetes mellitus in Depok. Offline data collection in 3 health centers in Depok using self-awareness questionnaire on the risk of stroke complications developed by itself with an alpha cronbach value of 0.826. The results of this study showed the majority were in the age range of 61-80 years, the gender of women and the majority of respondents adhered to Islam. The majority of respondents attended the last high school level, never attended DM counseling/education and have held DM for <5 years. The majority of respondents had self-awareness of the risk of moderate stroke complications. Respondents had a low level of self-awareness, more than respondents who had high self-awareness of the risk of stroke complications. This study recommends providing health education related to the risk of stroke complications in people with type 2 diabetes mellitus in order to increase self-awareness of the risk of stroke complications."

"Background: Platelet-Rich Plasma is believed to repair cartilage degeneration by stimulating tissue regeneration, however the clinical evidence is still insufficient. The aim of this research is to compare the efficacy of intra-articular hyaluronic acid injection (HA) and combined hyaluronic acid and platelet-rich plasma (HA-PRP) on osteoarthritis of the knee, and the association on gender, age, body mass index and osteoarthritis degree towards the efficacy. Methods: This is a cross-setional study using 58 patients’ medical records. 27 patients received HA injections, and 31 patients received HA-PRP injections. Patients were evaluated weekly using IKDC and WOMAC score until 2 months after the injections. Results: On the 2nd month of follow up, AH-PRP shows greater increase in IKDC (p=0,146) and WOMAC (Pain p=0,004; Stiffness p=0,008; Knee function p=0,007) score compared to HA injection. There’s no association on gender, age, and body mass index towards the IKDC and WOMAC score increase on either HA or combined HA-PRP injection. There’s no significant difference on osteoarthritis degree towards IKDC and WOMAC score increase on HA injection. However, there’s a significant difference between osteoarthritis degree 2 and 4 towards IKDC (p=0,002) and WOMAC score (Pain p=0,042; Stiffness p=0,018; Knee function p=0,042) and between osteoarthritis degree 2 and 3 towards WOMAC Pain (p=0,008) and WOMAC Knee function (p=0,018). Conclusions: HA-PRP injection shows better results to reduce osteoarthritis symptoms compared to HA injections. Better results were observed in patients with less severe osteoarthritis degree."

"Xenoprotein yang terkandung dalam medium ekspansi standar yang digunakan untuk kultur sel punca hematopoietik (SPH) CD34+ berisiko menyebabkan graft-versus-host disease pada pasien penerima cangkok SPH CD34+. Diperlukan suplementasi medium ekspansi xeno-free untuk menurunkan risiko graft-versus-host disease pada pasien penerima cangkok. Suplementasi medium kultur ekspansi menggunakan platelet-rich plasma (PRP) dan human serum albumin (HSA) yang keduanya berasal dari manusia diharapkan dapat menggantikan suplementasi xenoprotein dalam kultur. Platelet-rich plasma diketahui mampu meningkatkan laju proliferasi sel punca, sementara human serum albumin mampu mempertahankan kepuncaan sel punca lebih baik dari fetal bovine serum. Kombinasi PRP dan HSA sebagai suplementasi medium ekspansi diharapkan mampu meningkatkan proliferasi dan mempertahankan kepuncaan SPH CD34+. Pengaruh kombinasi PRP dan HSA, rasio optimal persentase gradien suplementasi PRP dan HSA, serta durasi optimal kultur yang mampu mendukung proliferasi dan mempertahankan sifat kepuncaan SPH CD34+ perlu diketahui. Jumlah sel hidup dihitung menggunakan metode eksklusi trypan blue untuk melihat kemampuan medium uji dalam mendukung proliferasi. Fenotipe SPH CD34+ dianalisis menggunakan flow cytometry untuk mengetahui kemampuan medium uji dalam mempertahankan kepuncaan. Kombinasi suplementasi PRP dan HSA mampu meningkatkan proliferasi dan mempertahankan kepuncaan hingga hari ke-7. Persentase gradien PRP : HSA terbaik merupakan 3 : 2 berdasarkan kemampuannya dalam meningkatkan proliferasi dan mempertahankan sifat kepuncaan SPH CD34+. Kombinasi PRP dan HSA memiliki efek positif terhadap kultur SPH CD34+

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Xenoprotein contained in CD34+ hematopoietic stem cell standard culture expansion medium has the risk of causing graft-versus-host disease (GVHD) in recipient of CD34+ HSC graft. Xeno-free supplementation in expansion medium is required to reduce the risk of GVHD in graft recipient. Supplementation of expansion medium using platelet-rich plasma (PRP) and human serum albumin (HSA), both originate from humans, hopefully has the ability to replace xenoprotein supplementation in culture. Platelet-rich plasma is known to increase the rate of stem cell proliferation, while human serum albumin is able to maintain stem cell’s stemness better than fetal bovine serum. The combination of PRP and HSA as expansion medium supplementation is expected to increase proliferation and maintain the stemness of CD34+ HSC. The effect of PRP and HAS combination, the optimal ratio of the percentage gradient of PRP and HSA supplementation, as well as the optimal duration of culture that can support proliferation and maintain CD34+ HSC stemness are to be studied. Live cells were counted using the trypan blue exclusion method to see the ability of the test medium to support proliferation. CD34+ HSC phenotype was analyzed using flow cytometry to determine the ability of test medium to maintain stemness. Combination of PRP and HSA supplementation are able to increase proliferation and maintain peaks until the 7th day. The best PRP : HSA gradient percentage is 3 : 2 based on its ability to increase proliferation and maintain SPH CD34+ stem properties. PRP and HSA combination has positive effects on CD34+ HSC culture."

"Diabetes melitus (DM) menyebabkan gangguan saraf autonom, sensorik, dan motorik, terutama di kaki dan pergelangan kaki yang menyebabkan perubahan postur kaki dan deformitas. Artritis pergelangan kaki penyandang DM mengakibatkan gangguan fungsional sehingga artrodesis merupakan suatu opsi tata laksana pembedahan. Sayangnya, gangguan metabolik DM mengakibatkan komplikasi yang tak jarang yakni non-union. Terapi sel dan derivatnya merupakan opsi terapi regeneratif untuk meningkatkan kesembuhan tulang.Pembuatan model artritis hewan DM dengan injeksi streptozotosin (STZ), diet tinggi lemak (HFD), dan injeksi complete freud adjuvant (CFA) dilanjutkan studi eksperimental in vivo dengan fokus hasil augmentasi fusi dengan granul hidroksiapatit (HA), sel punca mesenkimal asal tali pusat (SPM-TP), dan sekretom jaringan adiposa (JA). Sebanyak 16 sampel kaki dibagi menjadi 3 kelompok: kontrol negatif (kelompok 1), kontrol positif yang diberikan autologous bone graft (kelompok 2), dan perlakuan yang diberikan HA, SPM-TP, dan sekretom-JA (kelompok 3). Dilakukan evaluasi parameter klinis, radiologis, profil histomorfometris, dan ekspresi biomarker di subjek model artritis DM tikus Sprague Dawley (SD).Status DM tercapai setelah induksi STZ dengan rerata kadar glukosa 421 ± 27,16 mg/dL. Kelompok artritis menunjukkan perubahan diameter ankle yang bermakna dibanding kontrol serta perubahan radiologis sendi pergelangan kaki. Artritis berat (skor 3) ditemukan di mayoritas (80%) sampel kelompok 1 yang merupakan kontrol negatif (hanya induksi DM). Kelompok perlakuan menunjukkan skor artritis terendah serta osifikasi di sisi anterior tibiotalar. Terdapat perbedaan bermakna osteokalsin (p = 0,017) dan gen chordin (p = 0,003) antara ketiga kelompok.Simpulan: Model artritis DM pada tikus SD berhasil dibuat dengan injeksi STZ dan HFD serta induksi artritis kronik dengan injeksi CFA di sendi pergelangan kaki selama 4 minggu. Pemberian SPM-TP, sekretom JA, dan granul HA menunjukkan skor artritis yang lebih rendah. Namun, pemberian ketiga bahan tersebut tidak menghasilkan gambaran fusi yang lebih baik, serta tidak meningkatkan kadar osteokalsin, namun menghasilkan jumlah chordin (protein inhibisi BMP) yang lebih kecil dibandingkan baku standar.

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Diabetes mellitus (DM) can cause disturbances in the autonomic, sensory, and motor nerves, particularly in the feet and ankles, eventually leading to changes in foot posture and specific deformities. The advancement of management using stem cells and their secretome has shown promising outcomes. A model of DM arthritis can be created by inducing experimental animals with streptozotocin (STZ). In the DM arthritis model, the evaluation of ankle arthrodesis augmented with umbilical cord mesenchymal stem cell (MSC-Uc) and adipose tissue secretome (Secretome-AD) can be carried out to observe the existing outcomes.

This study is divided into two stages wherein the first stage involves creating an arthritis model in DM animals. The study utilizes a pretest-posttest design to measure clinical and laboratory parameters before and after treatment. Subsequently, the second stage involves an in vivo experimental study focusing on the outcomes of fusion augmentation with MSC-Uc, secretom-AD, and HA granule. The second stage of the study includes assessments using single-blinding methods for clinical, radiological, histomorphometric profile, and biomarker expression in the SD rat model of DM arthritis.

DM status was achieved after STZ injection, with an average glucose level of 421 ± 27.16 mg/dL. The final diameter averages in groups 1 – 3, which were not induced with arthritis (9.64 ± 0.49 mm), significantly differed from group 4 (12.50 ± 0.87 mm, p = 0.003) and 5 (11.85 ± 0.74 mm, p = 0.037). The arthritis groups showed radiological changes in the ankle joint. After modeling, there was a significant increase in fasting blood glucose compared to pre-modeling measurements. Severe arthritis (score 3) was found in the majority (80%) of samples in group 1, which served as the negative control (DM induction only). Anova test results for the IHC parameter showed significant differences (p = 0.017) in osteocalcin and chordin gene (p = 0.003) among the three groups.

Conclusion: A model of DM arthritis in SD rats was successfully created by STZ and HFD induction, followed by the induction of chronic arthritis with CFA injection in the ankle joint for 4 weeks. The administration of MSC-Uc, secretome-AD, and HA granule indicated lower arthritis scores. However, the administration of these three substances did not produce a better fusion picture, nor did it increase osteocalcin levels, but it resulted in a smaller amount of chordin (BMP inhibition protein) compared to the standard."