Background. Stromal vascular fraction (SVF) therapy has been performed over the past six years to treat 421 patients by our group in five clinical centers. Autologous SVF, which is a substance containing stem cells, was isolated from lipoaspirate, mixed with platelet-rich plasma (PRP), and administered to patients with degenerative diseases, autoimmune diseases, trauma, aging, and other diseases with unknown etiology. This study aimed to determine the safety of SVF and PRP that were given through infusion, spinal, and intra-articular injection. Methods. The lipoaspirate was treated with a tissue-dissociating enzyme, and then, through centrifugation, SVF was isolated. In addition, blood was drawn from each patient, and PRP was isolated. Autologous PRP and SVF were administered to all subjects by intravenous (IV) injection. A minority group within the population received an additional spinal or intra-articular injection. The type of intervention was determined by each disease evaluation. The cell doses and adverse events for each patient were documented and analyzed. Results. Cell dose that was considered to be safe was less than 10 billion SVF cells in 250 cc of normal saline, for IV injection, and less than 1 billion SVF, for intra-articular and spinal injection. Adverse events were not severe and were treated successfully. Any observed adverse events were identified as a result of spinal or intra-articular injections and were not related to SVF or PRP. Conclusions. Our results showed that administration of high dose of SVF until 10 billion cells in a majority of 421 patients through infusion, spinal, and intra-articular injection was feasible without causing major adverse events and should be further investigated in well-designed phase I-II clinical trial to address the safety and efficacy of therapy.

Background: Platelet-rich plasma (PRP) contains pro-angiogenic growth factors including vascular endothelial growth factor (VEGF). Angiogenesis is a necessary component of wound healing in instances of diabetic foot ulcers (DFU). PRP composition varies depending on methods and donor health status. Our group has developed an improved PRP protocol for diabetes treatment. The aims of this study were to examine the levels of the pro-angiogenic factor VEGF in these patient populations with and without diabetes.Methods: PRP was prepared using 24 mL of whole blood from 13 diabetic and 10 non-diabetic patients registered at Klinik Hayandra. Whole blood in sodium citrate tubes were centrifuged at 1,000 rpm for 5 minutes followed by plasma separation. Plasma samples were centrifuged at 3,000 rpm for 5 minutes.Upper platelet-poor plasma layers were discarded, leaving 5 mL of concentrated platelet containing plasma (PRP). Concentrated plasma samples were mixed, aliquoted, stored at −86 ℃, and pooled for platelet count, VEGF, and total protein analyses. Platelet counting was also performed using fresh whole blood and PRP to measure changes following PRP preparation.Results: Diabetic donors had higher whole blood platelet counts than non-diabetic donors, but this difference was not statistically significant. An average increase of more than 250% in platelet number after PRP preparation using our method was noted in both groups. Freezing-thawing samples at −86 ℃ lysed more than 90% of PRP platelets regardless of diabetes status. Diabetic PRP had lower mean total protein and higher VEGF concentrations. Lysed platelets from diabetic donors released more VEGF than those from non-diabetic donors.Conclusions: PRP from diabetic donors had higher VEGF content making autologous PRP application a promising treatment for DFU. However, this should be investigated another appropriate clinical trial.

Background: Ascorbic acid-2-phosphate has been reported to play a role in cell division and to suppress aging of cell. However, post-thawed cell morphology on various concentration of ascorbic acid is still unclear. In this study, we aimed to observe the morphology of post-thawed adipose-derived stem cells (ADSCs) in medium containing L-ascorbic acid-2-phosphate (LAA2P) (50 and 100 µg/mL).Methods: The cells were isolated from adipose tissue. Isolated cells then cultured and cryopreserved in liquid nitrogen. We detected mRNA expression of type 1 collagen on day 5. Cell seeded in T25 flask using basal medium [Dulbecco's modified Eagle's medium (DMEM) only] as a control group, DMEM with 10% fetal bovine serum (FBS) and antibiotics as DMFA group, while DMFA with ascorbic acid (50 and 100 µg/mL) as ascorbic acid treatment group. Results:The results showed that the cells cultured in DMEM only attached until 96 hours of observation while serum groups with or without ascorbic acid supplementation showed the proliferation until 240 hours of observation. The highest spread size of cell was in a serum group without ascorbic acid supplementation and the highest yield of cells showed in a group with 50 µg/mL of ascorbic acid supplementation. Reduced mRNA expression of type 1 collagen which related to aging was showed in cells cultured without ascorbic acid supplementation.Conclusions: These results showed that ascorbic acid increased the cell division and suppressed the aging processes indicated by normal spread cell in size compared to cell cultured in DMFA without ascorbic acid supplementation.

AbstrakStudi tentang kemampuan adipose-derived stem cells (ADSCs) sebagai sel punca yang dapat berdiferensiasi menjadi kondrosit menggunakan platelet-rich plasma (PRP) sebagai subtitusi fetal bovine serum (FBS) telah banyak dilaporkan. Penggunaan medium pertumbuhan dengan kombinasi ADSCs, PRP dan scaffold sutra masih belum banyak dipelajari dalam rekayasa jaringan kartilago. Studi ini bertujuan untuk mempelajari pengaruh medium yang mengandung 5%, 10% dan 20% PRP terhadap proses kondrogenesis ADSCs manusia yang dikultur pada scaffold sutra Bombyx mori Indonesia. Metode penelitian diawali dengan pembuatan scaffold sutra menggunakan metode salt-leaching, isolasi dan kultur ADSCs manusia dari jaringan lemak, uji pertumbuhan ADSCs pada scaffold sutra dengan variasi konsentrasi PRP pada medium serta analisis kadar glikosaminoglikan (GAG). Hasil penelitian menunjukkan bahwa ADSCs yang dikultur menggunakan PRP lebih tinggi laju pertumbuhannya dibandingkan dikultur menggunakan FBS selama 7 hari pengamatan. Kadar GAG yang disekresikan ADSCs kelompok PRP juga lebih tinggi dibandingkan kelompok FBS. Kadar GAG tertinggi pada hari ke-21 pengamatan adalah medium yang mengandung 20% PRP kemudian 10% dan 5%, sedangkan kadar GAG kelompok kontrol cenderung stabil pada kadar yang rendah. Berdasarkan hasil tersebut, medium yang mengandung PRP memiliki potensi dalam menginduksi kondrogenesis ADSCs yang dikultur pada scaffold sutra.Abstract The studies on adipose-derived stem cells (ADSCs) differentiation into chondrocytes using platelet-rich plasma (PRP) as a substitute for fetal bovine serum (FBS) have been reported. However, the combination of ADSCs, PRP and silk fibroin scaffold has not been widely studied for developing cartilage engineering. Therefore, this research aims to study the effect of medium containing 5%, 10% and 20% PRP towards chondrogenesis of human ADSCs cultured on silk fibroin scaffold from Indonesia Bombyx mori. At first, the silk fibroin scaffold was fabricated using a salt-leaching method, then ADSCs were isolated and cultured from adipose tissues. The assays of growth curve and biocompatibility of silk fibroin scaffold toward ADSCs supplemented by PRP as well as glycosaminoglycans (GAG) concentration were conducted later. The results showed that higher absorbance of proliferation rate was on ADSCs supplemented by various PRP concentrations compare to FBS control group for seven days of observation. Level of GAG, which secreted by ADSCs supplemented by a various concentration of PRP, was also higher than the FBS group. The highest level of GAG on day 21 was observed in 20% PRP group then 10% and 5% PRP, while a group of GAG level is stable at low levels. This study concludes that PRP has the potential to induce chondrogenesis ADSCs which cultured on silk fibroin scaffold.

No abstract

Defisiensi vitamin D masih menjadi masalah kesehatan global sampai saat ini, tidak terkecuali negara tropis yang memiliki lebih banyak paparan sinar matahari. Studi di negara-negara Eropa, Amerika dan Asia menunjukkan tingkat defisiensi vitamin D berkisar antara 45-90%. Indonesia termasuk negara dengan tingkat defisiensi vitamin D yang tinggi, tetapi laporan mengenai status vitamin D dan upaya pencegahannya belum banyak dilakukan. Keluhan pada muskuloskeletal akan menimbulkan gangguan atau sensasi ketidaknyamanan, memengaruhi performa dalam bekerja dan mengurangi produktivitas terutama pada kelompok usia dewasa. Penelitian ini bertujuan untuk mengetahui hubungan antara asupan vitamin D dan keluhan muskuloskeletal pada kelompok usia 17-35 tahun pada mahasuswa Fakultas Kedokteran dan Ilmu Kesehatan (FKIK) Unika Atma Jaya dan kerabatnya pada rentang usia tersebut. Desain penelitian observasional analitik dengan pendekatan potong lintang. Pengambilan data dilakukan dengan metode consecutive sampling pada tahun 2021 di Indonesia. Asupan vitamin D diukur dengan Vitamin D Estimation Only - Food Frequency Questionnaire dan keluhan muskuloskeletal diukur dengan kuesioner Nordic Body Map. Data dianalisis dengan menggunakan uji Chi-square. Subjek penelitian sebanyak 108 responden. Hasil penelitian menunjukkan terdapat 67,6% responden dengan asupan vitamin D yang tidak adekuat dan 80,6% yang memiliki keluhan muskuloskeletal. Tidak terdapat hubungan yang signifikan antara asupan vitamin D dengan keluhan muskuloskeletal pada kelompok usia 17-35 tahun (P-value= 0,379).

Burns increase tissue damage by free radicals, thus hindering the healing process. Platelet Rich Fibrin (PRF) and Advanced Platelet Rich Fibrin (A-PRF) are autologous fibrin matrices that contain growth factors and have been proven to increase tissue formation and regeneration. Objectives: To determined the effect of applying PRF and A-PRF on deep second-degree burns. Methods: This study was experimental research on twenty male Sprague Dawley rats were randomly divided into four groups. Group A and B as control groups received NaCl 0,9% and silver sulfadiazine ointment, respectively. Group C was given PRF and group D was given A-PRF. Burns are photographed and the area was calculated using the ImageJ application. After 14 days of therapy, the burn areas were excised and made into histological slides. Microscopic study of angiogenesis was observed using a microscope at 400x magnification. Results: Changes in wound surface area in PRF and A-PRF groups were 3,106 cm and 2,333 cm. The number of angiogenesis in PRF and A-PRF groups were 3.88 and 5.88. The data was analyzed using One-Way Anova and the results showed that there were no significant differences between the changes in the wound area and angiogenesis of PRF and A-PRF groups. Conclusion: This study showed that there are no significant differences in changes in wound surface area and angiogenesis of burn injury after topical application of PRF and A-PRF.Keywords:  angiogenesis, burn injury, changes in wound surface area, A-PRF, PRF

Angiogenesis merupakan proses penting untuk pertumbuhan dan regenerasi sel. Salah satu protein yang berperan dalam angiogenesis adalah vascular endothelial growth factor (VEGF).  VEGF akan berinteraksi dengan reseptornya seperti vascular endothelial growth factor receptor-2 (VEGFR2) untuk meregulasi proses angiogenesis. Mutasi pada gen VEGFR2 pada manusia dilaporkan dapat menyebabkan penyakit strawberry mark (hemangioma). Oleh karena itu, perlu dilakukan studi dan penelusuran terkait mutasi yang berperan dalam terbentuknya hemangioma. Beberapa tahapan yang dilakukan untuk menganalisis pengaruh mutasi tersebut diantaranya dengan melakukan penelusuran informasi dasar dan komposisi gen VEGFR2, analisis ortolog dan paralog gen VEGFR2, dan analisis struktur sekunder protein VEGFR2 manusia. Hasilnya menunjukkan bahwa mutasi VEGFR2 pada basa ke-3741 yaitu sitosin (C) menjadi timin (T) dilaporkan ditemukan pada jaringan tumor hemangioma. Perubahan basa ke-3741 sitosin menjadi timin mengakibatkan perubahan asam amino prolin menjadi serin yang dapat berdampak pada regulasi ekspresi VEGF. Ortolog dan paralog dari gen VEGFR2 manusia adalah gen VEGFR2 mencit dan gen fibroblast growth factor receptor 1 (FGFR1) manusia. Hasil analisis pengaruh mutasi terhadap bentuk dan struktur protein menunjukkan tidak ada perubahan yang signifikan, namun posisi mutasi di bagian ekstraseluler sel diduga mempengaruhi regulasi ekspresi VEGF melalui ikatan ligan-reseptor.