Comparative in vitro study of the cytotoxicity of gelatine and alginate to human umbilical cord mesenchymal stem cells

Background: Mesenchymal stem cells (MSCs) and scaffold combination constitute a promising approach currently adopted for tissue engineering. Umbilical cord-derived mesenchymal stem cells (hUC-MSCs) are easily obtained and non-invasive. Gelatine and alginate constitute a biocompatible natural polymer scaffold. At present, a cytotoxicity comparison of gelatine and alginate to hUC-MSCs is not widely conducted. Purpose: This study aimed to compare the cytotoxicity of gelatine and alginate in hUC-MSCs in vitro. Methods: Isolation and culture were performed on hUC-MSCs derived from healthy full-term neonates. Flow Cytometry CD90, CD105 and CD73 phenotype characterization was performed in passage 4. 3-(4,5-dimethythiazol- 2-yl)-2,5-diphenyl tetrazolium bromide (MTT) colorimetric assay was performed to measure the cytotoxicity. The three sample groups were: (T1) hUC-MSCs with α-MEM (alphaminimum essential medium) solution as control; (T2) hUC-MSCs with gelatine; (T3) hUC-MSCs with alginate. Results: Flow cytometry of hUC-MSCs displayed positive CD90, CD105 and CD73 surface markers. Gelatine and alginate had no effect on the viability of hUC-MSCs and no statistically significant difference (p>0.05) of cytotoxicity between gelatine and alginate to hUC-MSCs. Conclusion: Gelatine and alginate proved to be non-toxic to hUC-MSCs in vitro

Studio Biocompatibility and osteogenic differentiation potential of human umbilical cord mesenchmal stem cell (hUCMSCs) with gelatin solvet

Dengan ini dinyatakan dengan sebenarnya bahwa karya ilmiah ini telah diperiksa/divalidasi dan hasilnya telah memenuhi kaidah ilmiah, norma akademik dan norma hukum sesuai dengan peraturan menteri pendidikan nasional 17 tahun2010

A Potential therapy of human umbilical cord mesenchymal stecells for bone regeneration on osteoporotic mandibular bone

Dengan ini di nyatakan dengan sebenarnya bahwa karya ilmiah ini telah diperiksa / divalidasi dan hasilnya telah memenuhi kaidah ilmiah, norma akademika dan norma hukum sesuai dengan peraturan menteri pendidikan nasional 17 tahun 2010

Phenotype Characteristics and Osteogenic Differentiation Potential of Human Mesenchymal Stem Cells Derived from Amnion Membrane (HAMSCs) and Umbilical Cord (HUC-MSCs)

Introduction: Human amnion membrane mesenchymal stem cells (hAMSCs) and human umbilical cord mesenchymal stem cells (hUC-MSCs) are potential, non invasive sources of stem cells used for bone tissue engineering. Phenotyping characterization is an extremely important consideration in the choice of the appropriate passage in order to maximize its osteogenic differentiation potential. Aim: To explore phenotype characteristics and compare osteogenic differentiation potential of hAMSCs and hUC-MSCs. Method: Isolation and culture were performed on hAMSCs and hUC-MSCs from a healthy woman in her 38th weeks of pregnancy. CD90, CD105 and CD73 phenotype characterization was done in passage 4-7. An osteogenic differentiation examination of hAMSCs and hUC-MSCs with Alizarin red staining and RUNX2 expression was performed in the passage that had appropriate expressions of phenotype characteristics. Results: The expression of CD90 hUC-MSCs was higher than that of hAMSCs in all passages. CD105 hUC-MSCs was higher in passage 4-6, while CD105 hAMSCs was equal to that of hUC-MSCs in passage 7. CD73 hUC-MSCs was higher than hAMSCs in passage 4 and 5, while in passage 6 and 7 hAMSCs was higher than hUC-MSCs. There was a decrease in the number of CD90, CD105 and CD73 on hAMSCs and hUC-MSCs in passage 5, then determined as appropriate passage. Alizarin red staining examination showed calcium deposition and revealed no significant difference, but RUNX2 expression of hUC-MSCs was significantly higher than that for hAMSCs. Conclusion: Both hAMSCs and hUC-MSCs had phenotype characteristics of mesenchymal stem cell and showed ostegenic differentiation potential

Immediate Complete Overdenture for Esthetic Consideration of the Upper Edentulous Ridge - A Case Report

Background: Nowadays esthetics is one of the main considerations. So, dentures not only rehabilitate functions, but esthetics as well. The problem is that “conventional denture making” requires several steps which sometimes force a patient into a toothless phase which affects both functional and esthetical aspects. In some cases, the patients are unable to endure this condition due to esthetic reasons. Immediate dentures will be the best solution for them. Purpose: To provide information about immediate complete overdenture design and fabrication. Case: A 61-year-old female patient, an active physician, came to the Dental Hospital of Airlangga University, complaining about her upper partial denture and lower complete denture, which had been worn for 10 years. They weren’t comfortable to wear anymore. She wanted new dentures, without the toothless phase. The remaining teeth were 16, 13, 12, and 22. 13 and 22 were used as retainers for the upper partial denture but the inclination and vitality of the teeth were bad. Case management: Immediate complete overdenture was made for the upper edentulous ridge. Immediate treatment was chosen because of the esthetic consideration. Overdenture treatment was chosen because it preserves the bone dimensions. 16 and 12 were extracted, while 13 and 22 were endodontically treated. The complete denture was then fabricated by cutting of 13 and 22 on the functional model. After the complete denture had been fabricated, the patient’s teeth 13 and 22 were decaputated as high as the gingival level and then restored with GIC on the surface. The denture was then inserted immediately after the decaputations. Discussion: An immediate denture is a denture which is made first before decaputation or extraction, then immediately inserted into the mouth after the tooth decaputation or extraction. Designing an immediate denture will give some advantages to patients, especially from the esthetic aspect. An overdenture is a good treatment for patients whose condition of teeth crowns is bad but periodontal tissue and teeth roots are in good condition. In conclusion, an immediate denture is the best treatment where esthetics is the main consideratio

Human Umbilical Cord Mesenchymal Stem-Cell Therapy to Increase the Density of Osteoporotic Mandibular Bone

Objective The aim of this study is to evaluate the feasibility of human umbilical cord mesenchymal stem-cell (hUCMSC) therapy in increasing osteoporotic mandibular bone density in a rat model by determining changes in alkaline phosphatase (ALP), osteocalcin, type 1 collagen, and trabecular bone area after treatment. Materials and Methods This research adopted an experimental posttest-only control group design. Thirty female Wistar rats were randomly divided into six groups, namely, a control group with rats postsham surgery (T1), osteoporotic model postovariectomy rats (T2), postovariectomy rats 4 weeks after gelatin injection (T3), postovariectomy rats 8 weeks after gelatin injection (T4), postovariectomy rats 4 weeks after hUCMSC injection (T5), and postovariectomy rats 8 weeks after hUCMSC injection (T6). The rats were all sacrificed for histological and immunohistochemical examinations of ALP, osteocalcin, type 1 collagen, and trabecular bone area. Results Increased expression of ALP, type 1 collagen, and osteocalcin, as well as increased trabecular bone area, was observed in the treatment groups compared with that in the osteoporotic groups. Conclusion hUCMSCs produce significant osteogenic effects and increase osteoporotic mandibular bone density in the animal model. Increases in bone density are demonstrated by the higher levels of ALP, osteocalcin, and type 1 collagen, as well as increases in the trabecular bone area

The Effect of a combination of 12% spirulina and 20% chitosan on macrophage, PMN, and lymphocyte cell expressions in post extraction wound

Background: Tooth extraction is the ultimate treatment option for defective teeth followed by the need for dentures. Inflammation is one phase of the healing process that should be minimized in order to preserve alveolar bone for denture support. Macrophage, PMN and lymphocyte cells are indicators of acute inflammation. Spirulina and chitosan are natural compounds with the potential to be anti-inflammatory agents. Purpose: This study aimed to determine macrophage, PMN and lymphocyte cells of animal models treated with a combination of 12% spirulina and 20% chitosan on the 1st, 2nd and 3rd post-extraction day. Methods: Animal models were randomly divided into control (K) and treatment (P) groups. Each group was further divided into three subgroups (KI, KII, KIII and PI, PII, PIII). The post-extraction sockets of the control group animals were then filled with CMC Na 3%. Meanwhile, the post-extraction sockets of the treatment group members were filled with a combination of 12% spirulina and 20% chitosan. Subsequently, the number of PMN, macrophage and lymphocyte cells was analyzed by means of HE analysis on the 1st, 2nd and 3rd days. Statistical analysis was then performed using a T-test. Results: There was a decrease in PMN cells and an increase in macrophage and lymphocyte cells on Days 1, 2 and 3. Conclusion: It can be concluded that a combination of 12% spirulina and 20% chitosan can not only decrease PMN cells, but can also increase macrophage and lymphocyte cells on day 1, 2 and 3 after tooth extraction

The Correlation of Bone Mineral Density (BMD), Body Mass Index (BMI) and Osteocalcin in Postmenopausal Women

Background: Osteoporosis is related to the decrease of bone density, osteoporosis requires attention from dentists because it can also occurs in the jaw bone. Osteoporosis is calculated with a quantitative assessment of bone density, which is called Bone Mineral Density (BMD). The best imaging modalities to assess bone density are the Dual Energy examination methods X-ray Absorptiometry (DEXA). In addition to DEXA, osteoporosis examination can be performed with measurement of Body Mass Index (BMI), which is a ratio of weight and height. Process of bone formation by osteoblasts can be examined by bone marker such as Osteocalcin, as a parameter (alone or in combination with BMD) to determine metabolic bone disorders during bone formation and bone remodeling (bone turn over). Aim: To analyze the correlation among BMD, BMI and Osteocalcin in Postmenopausal women, to investigate the possibility of using osteocalcin examination to predict the mandibular bone osteoporosis. Method: Fifty four women above 51 years old who are at least 1 year postmenopausal, underwent BMD (using DEXA examinations), BMI, and Osteocalcin. The results of each examination were counted, and the correlations among each examinations were evaluated using Spearman’s correlation test. Result: Means of BMD, BMI, and Osteocalcin in postmenopausal women were 1.606, 25.189, and 30.566 respectively. BMD were significantly correlated with BMI (Spearman’s rank correlation coefficient r=0.414, p<0.05). While, BMD were significantly correlated with Osteocalcin (r=-0.343, p<0.05). Moreover, BMI were significantly correlated with Osteocalcin (r=-0.274, p<0.05). Conclusion: There were significant correlations among BMD, BMI, and Osteocalcin results in postmenopausal women. It is concluded that each examination of BMD, BMI, and Osteocalcin can be used to identify the risk of osteoporosis in postmenopausal women. Therefore simple examination of osteocalcin can be used to predict mandibular bone loss

A potential therapy of human umbilical cord mesenchymal stem cells for bone regeneration on osteoporotic mandibular bone

Objective: The aim of this study is to prove that human umbilical cord mesenchymal stem cell (hUCMSC) therapy on mandibular osteoporotic model is able to increase transforming growth factor‑beta‑1 (TGF)‑β1 expression, Runx2, and osteoblasts. Materials and Methods: This research is true experimental posttest control group design. Thirty female Wistar rats were divided into 6 groups randomly, which consisted of sham surgery for control (T1), ovariectomy as osteoporotic group (T2), osteoporotic group injected with gelatine for 4 weeks (T3), 8 weeks (T4) injected with hUCMSC‑gelatine for 4 weeks (T5) and 8 weeks (T6). All mice were presented for immunohistochemistry examination for TGF‑β1, Runx2, and histology for osteoblasts. Results: The lowest level of osteoblast was osteoporotic group injected with gelatine in 4 weeks compared to other groups. There were increases of TGF‑β1, Runx2, and osteoblasts from osteoporotic group compared to osteoporotic post‑hUCMSC‑gelatine injection group. Conclusion: The hUCMSC has a high osteogenic effect and increases the osteoporotic mandibular bone regeneration on the animal model that is showed by the increase of the level of TGF‑β1, Runx2, and osteoblast

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