Fibrin Glue (FG) Attenuates Fibrosis On Human Tenon's Fibroblasts (HYFS) of Glaucomatous Eyes : Comparison with Mitomycin C

Human tenon’s fibroblasts (HTFs) are major cells that contribute to the fibrotic response that usually occurs after trabeculectomy. The activation of HTFs into myofibroblast contributes to the disturbance of the extracellular matrix (ECM) remodeling. Excessive ECM deposition, mainly collagen type I, with cell contraction is the major hallmark of bleb fibrosis. HTFs were isolated from the tenon’s tissue of a glaucomatous patient. HTFs were divided into 3 groups which consisted of FBS 2% control group, MMC and FG treated group. This study investigated the effects of FG on cell viability, cell migration, cell contractility, collagen synthesis and degradation in HTFs. Cell viability was determined by MTT-assay, while collagen synthesis and degradation were determined by Sirius red binding assay. Cell migration was carried out by in vitro scratch assay, and cell contractility was analyzed by fibroblast populated-collagen gel assay. The differences in cell viability, cell contractility, collagen synthesis and degradation among the various groups were analyzed using One-way ANOVA or Kruskal Wallis test, followed by a posthoc test with 95% confidence interval (p<0.05). FG significantly decreased collagen synthesis (31.31±2.91ug/mL) in line with the induction of collagen degradation (85.50±3.16ug/mL) in HTFs when compared to FBS 2% control. While FG inhibits cell contraction while maintaining cell viability in HTFs, MMC provides a better antifibrotic effect in HTFs. FG may inhibit fibrosis formation of HTFs by inhibiting collagen synthesis and cell contraction while inducing collagen degradation. FG might have an antifibrotic effect on HTFs through extracellular matrix remodeling

The Role of Stem Cell Metabolites Derived From Placenta for Skin Regeneration An In Vitro Study

Background: The role of stem cells in skin aging is to repair injured tissue or replace other cells in programmed cell death. Stem cell metabolites are rich in growth factors including IL-10, IL-4, EGF, GM-CSF, and TGF-β that can induce the skin production of protein and elastic fbers, leading to the improvement of skin appearance. Aim: This study aimed to assess the characteristics of stem cell metabolites in vitro. Methods: Cytotoxicity assay was performed using MTT reagents and optical cell densities were determined using ELISA reader to fnd the percentage of living cells. Cytokine detection assay was performed by analyzing the cytokine levels in the peripheral blood mononuclear cell (PBMC) and mesenchymal stem cell (MSC) using ELISA. Apoptosis assay was performed using the double staining method with the markers identifed were Hsp70, p53, and caspase-3. Results: All samples showed the percentage of living cells that exceed 70%. Cytokine detection assay showed a decrease of IL- 12 and IFN-γ in both PBMC and MSC groups. The apoptosis assay of human adipose mesenchymal stem cells using a fluorescence microscope showed most of the green light was lost in control cells without metabolites. We found that the expressions of Hsp70 were increased while the expression of p53 and caspase-3 were decreased in the stem cell metabolites samples. Conclusion: These results showed that stem cell metabolites are non-toxic, do not cause a systemic immune response to surrounding tissue, and able to inhibit the occurrence of apoptosis

Pre-Clinical Trial Stem Cell Metabolites Derived From Placenta For Wound Healing

Previous research focuses on in vitro study of stem cell metabolites derived from placenta for wound healing. This study, however, is an advanced stage which focuses on testing the efficiency and efficacy of stem cell metabolites in rats (Rattus novergicus). The tests carried out examined the blood levels with ELISA instruments and integument histology by observing the activity of polymorph nuclear and monocyte cells in the control and treatment groups. In the control group, the rats were injured in the anterior and posterior back skin with a 1×1 cm incision wound, (only antibiotics), while the treatment group uses antibiotics and 4 mL injections of stem cell metabolites. Each group was repeated three times with the samples observed for blood levels using ELISA Interleukin-4, Interleukin-10 and Tumor Necrosis Factor-α, with integument histology at pre-injection, in days 1, 3 and 6. These were used to compare the development of inflammatory cells, polymorphonuclear and monocytes between the control and treatment groups. Stem cell metabolites are significantly effective and efficient with the ability to inhibit the inflammatory process in tissues in terms of examining the blood levels of rats using ELISA Interleukin-4, Interleukin-10 and Tumor Necrosis Factor-α. Interleukin-4 and Interleukin-10 (anti-inflammatory) tend to significantly increase the treatment group, while Tumor Necrosis Factor-α (pro inflammation) increases the control group. Histology results showed a decrease in the activity of polymorphonuclear and monocytes inflammatory cells in the treatment group compared to the control, which indicated that the stem cell metabolites were able to significantly inhibit the inflammatory process. It is concluded that stem cell metabolites derived from placenta are effective and efficient for wound healing in rats. Clinical study is needed for further research for it to be used on humans

Pre-Clinical Trial Stem Cell Metabolites Derived from Placenta for Wound Healing

Previous research focuses on in vitro study of stem cell metabolites derived from placenta for wound healing. This study, however, is an advanced stage which focuses on testing the efficiency and efficacy of stem cell metabolites in rats (Rattus novergicus). The tests carried out examined the blood levels with ELISA instruments and integument histology by observing the activity of polymorph nuclear and monocyte cells in the control and treatment groups. In the control group, the rats were injured in the anterior and posterior back skin with a 1×1 cm incision wound, (only antibiotics), while the treatment group uses antibiotics and 4 mL injections of stem cell metabolites. Each group was repeated three times with the samples observed for blood levels using ELISAInterleukin-4, Interleukin-10 and Tumor Necrosis Factor-α, with integument histology at pre-injection, in days 1, 3 and 6. These were used to compare the development of inflammatory cells, polymorphonuclear and monocytes between the control and treatment groups. Stem cell metabolites are significantly effective and efficient with the ability to inhibit the inflammatory process in tissues in terms of examining the blood levels of rats using ELISA Interleukin-4, Interleukin-10 and Tumor Necrosis Factor-α. Interleukin-4 and Interleukin-10 (anti-inflammatory) tend to significantly increase the treatment group, while Tumor Necrosis Factor-α (pro-inflammation) increases the control group. Histology results showed a decrease in the activity of polymorphonuclear and monocytes inflammatory cells in the treatment group compared to the control, which indicated that the stem cell metabolites were able to significantly inhibit the inflammatory process. It is concluded that stem cell metabolites derived from placenta are effective and efficient for wound healing in rats. Clinical study is needed for further research for it to be used on humans

Study of Human Amniotic Membrane Mesenchymal Stem Cells Using Gelatin and Alginate as Nontoxic Scaffolds

Perinatal mesenchymal stem cells (MSCs), for example, from amniotic membrane, have advantages over adult sources, such as bone marrow, in terms of ease of availability, cell naivety, tissue stem cell abundance, high capacity of proliferation, and less donor site morbidity, because it does not require invasive procedures. Natural polymer scaffolds, such as gelatin and alginate, are biocompatible. Combination of stem cells from amniotic membrane (hAMSCs) and gelatin or alginate as scaffold can be promising. However, cytotoxicity comparison of gelatin and alginate to hAMSCs has not been widely studied. This study was aimed to compare cytotoxicity of gelatin and alginate on hAMSCs in vitro. Isolation and culture were performed on hAMSCs of the healthy full-term pregnancy. In passage 4, Flow Cytometry CD90, CD105, and CD73 phenotype characterization was done. Colorimetric assay of 3-(4,5-dimethythiazol-2-yl)-2,5 diphenyl tetrazolium bromide (MTT) was performed to measure the cytotoxicity. There were three sample groups: (control group) hAMSCs with alpha-minimum essential medium (α-MEM) solution as control; (gelatin group) hAMSCs with gelatin; (alginate group) hAMSCs with alginate. Each group consisted of 12 samples. Flow cytometry of hAMSCs expressed 28.78% CD90, 36.95% CD105, and 44.41% CD73 surface markers. No sample depicted toxicity in either gelatin or alginate group, and this is indicated by the average percentage of living cells in gelatin 97.26% and in alginate 98.43%. No statistically significant difference (p=0.057) of cytotoxicity was found between gelatin and alginate to hAMSCs. Gelatin and alginate were nontoxic to hAMSCs in vitro

A Potential Differentiation of Adipose and Hair Follicle-derived Mesenchymal Stem Cells to Generate Neurons Induced with EGF, FGF, PDGF and Forskolin.

Human Adipose Derived Mesenchymal Stem Cells (HADMSCs) and Human Hair Follicle Derived Mesenchymal Stem Cells (HHFDMSCs) have attracted great interest because of their multilineage differentiation potential, selfrenewal properties, and their possible use of cell and gene therapies. This present study to investigate the neurogenic differentiation ability of hADMSCs and hHFDMSCs induced by Epidermal Growth Factors (EGF), Fibroblast Growth Factor (FGF), Platelet Derived Growth Factor (PDGF) and Forskolin. This study was true experimental with longitudinal study design. The sample size determined with minimal sample size formula and it was randomly chosen. These studies employed an in vitro design for the expansion and proliferation of Mesenchymal Stem Cells (MSCs) and examined the heterogeneity of these cells using the markers CD105, CD90, OCT4, and SOX2. MSCs from adipose tissue and hair follicles were induced with EGF, FGF, PDGF and Forskolin to differentiate and generate neurons. The capacity of MSCs to generate neurons were verified using glial fibrillary acidic protein, nestin, and β-tubulin III . The expression of neural markers and morphological changes in Mesenchymal stem cells from hADMSCs and hHFDMSCs were confirmed. hADMSCs and hHFDMSCs share a similar capacity to differentiate and generate neurons, which is beneficial for the development of neuronal restoration for future therapies for patients suffering from neurological diseases

Gingival Mesenchymal Stem Cells from Wistar Rat’s Gingiva (Rattus Novergicus) – Isolation and Characterization (In Vitro Study)

Gingiva is emerging as a source of Mesenchymal Stem Cells. Gingival Mesenchymal Stem Cell has been isolated and characterized from the gingival connective tissue of wistar rat (Rattus Novergicus). Gingival Mesenchymal Stem Cell sources are rich, attainable and easy to isolate through minimal invasive procedure. Gingival Mesenchymal Stem Cells are ideal to accelerate bone regeneration. The aim of this study was to analyze Gingival Mesenchymal Stem Cells from Wistar Rats’ gingiva (Rattus Novergicus) isolation and characterization by CD34, CD44, CD73, CD90, CD105 expression. This study was descriptive observational with simple random sampling method. Gingival Mesenchymal Stem Cells were isolated from healthy, 200 gram, 1 month year old, male rat’s (Rattus Novergicus) lower gingival tissue through gingivectomy procedure (n=4). Gingiva were minced into small fragments then cultured in 2 weeks. The culture was passaged every 3-5 days after cultured and plated. The isolated Gingival Mesenchymal Stem Cells in passage 5 were characterized by CD34, CD44, CD73, CD90, CD105 using Immunocytochemistry and flowcytometry examination. Gingival Mesenchymal Stem Cells strongly expressed CD44+, CD73+, CD90+, CD105+ but did not express CD45- and CD34-. Gingival Mesenchymal Stem Cells’ morphology was fibroblast-like, spindle-shaped, colony-forming abilities, and stick to the culture plate. Gingiva is potential Stem Cell source. Gingival Mesenchymal Stem Cells has multipotency ability with proliferation and mesenchymal stem cells characteristic advantageous for tissue engineering and regenerative therapy

THE CHANGING CLINICAL PERFORMANCE OF DENGUE VIRUS INFECTION IN THE YEAR 2009

Background: Dengue (DEN) virus, the most important arthropod-borne human pathogen, represents a serious public health threat. DEN virus is transmitted to humans by the bite of the domestic mosquito, Aedes aegypti, and circulates in nature as four distinct serological types DEN-1 to 4). The aim of Study: To identify Dengue Virus Serotype I which showed mild clinical performance in five years before and afterward showed severe clinical performance. Material and Method: Prospective and analytic observational study had been done in Dr. Soetomo Hospital and the ethical clearance was conduct on January 01, 2009. The population of this research is all cases of dengue virus infection. Diagnosis were done based on WHO 1997. All of these cases were examined for IgM & IgG anti Dengue Virus and then were followed by PCR examination to identify Dengue Virus serotype. Result and Discussion: DEN 2 was predominant virus serotype with produced a spectrum clinical illness from asymptomatic, mild illness to classic dengue fever (DF) to the most severe form of illness (DHF). But DEN 1 usually showed mild illness. Helen at al (2009–2010) epidemiologic study of Dengue Virus Infection in Health Centre Surabaya and Mother and Child Health Soerya Sidoarjo found many cases of Dengue Hemorrhagic Fever were caused by DEN 1 Genotype IV. Amor (2009) study in Dr. Soetomo Hospital found DEN 1 showed severe clinical performance of primary Dengue Virus Infection as Dengue Shock Syndrome two cases and one unusual case. Conclusion: The epidemiologic study of Dengue Virus Infection in Surabaya and Sidoarjo; in the year 2009 found changing predominant Dengue Virus Serotype from Dengue Virus II to Dengue Virus 1 Genotype IV which showed a severe clinical performance coincident with primary infection

Metode Pembuatan Metabolit Sel Punca Dari Darah Tali Pusat Manusia Untuk Regenerasi Kulit

Invensi ini berkaitan dengan metode pemrosesan produk metabolit stem cell untuk regenerasi kulit yang dilakukan di Laboratorium. Etabolit stem cell yang dihasilkan berasal dari darah tali pusat bayi dari honor sukarela yang memenuhi standar, untuk kemudian dilakukan kultur dan diferensiasi menjadi stem cell mesenkim. Kandungan sitokin dalam metabolit stem cell yaitu IL-10, IL-4, EGF, GM-CSF dan TGF-B yang berperan dalam mempercepat regenerasi kulit, merangsang pertumbuhan dan perkembangan berbagai sel kekebalan, menggantikan sel-sel yang rusak, membuat kulit memproduksi sendiri serat elastic protein agar kulit kembali elastis, serta memproduksi lebih banyak kolagen dan elastin. Darah tali pusat diisolasi kemudian dikultur pada komplit medium dalam sistem bioreaktor. Selanjutnya dikarakterisasi menggunakan flow cytometry dengan marker psoitif CD105 dan CD90