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1.
Article | IMSEAR | ID: sea-195873

ABSTRACT

Background & objectives: Seeding density is one of the major parameters affecting the quality of tissue-engineered cartilage. The objective of this study was to evaluate different seeding densities of osteoarthritis chondrocytes (OACs) to obtain the highest quality cartilage. Methods: The OACs were expanded from passage 0 (P0) to P3, and cells in each passage were analyzed for gross morphology, growth rate, RNA expression and immunochemistry (IHC). The harvested OACs were assigned into two groups: low (1×10[7] cells/ml) and high (3×10[7] cells/ml) cell density. Three-dimensional (3D) constructs for each group were created using polymerised fibrin and cultured for 7, 14 and 21 days in vitro using chondrocyte growth medium. OAC constructs were analyzed with gross assessments and microscopic evaluation using standard histology, IHC and immunofluorescence staining, in addition to gene expression and biochemical analyses to evaluate tissue development. Results: Constructs with a high seeding density of 3×10[7] cells/ml were associated with better quality cartilage-like tissue than those seeded with 1×10[7] cells/ml based on overall tissue formation, cell association and extracellular matrix distribution. The chondrogenic properties of the constructs were further confirmed by the expression of genes encoding aggrecan core protein and collagen type II. Interpretation & conclusions: Our results confirmed that cell density was a significant factor affecting cell behaviour and aggregate production, and this was important for establishing good quality cartilage.

2.
The Medical Journal of Malaysia ; : 121-122, 2008.
Article in Malayalam | WPRIM | ID: wpr-629999

ABSTRACT

Bone marrow derived progenitor cells have been widely studied for its multipotent property and have proofed to be an important resource in regenerative medicine. However, the propagation of murine bone marrow appeared to be a great challenge as compared to other mammalian species. In this study, various isolation techniques and the plasticity of the isolated cells were evaluated. Our result shows that magnetic sorting technique yielded the most viable cells and displayed wider differentiation capacity.

3.
The Medical Journal of Malaysia ; : 117-118, 2008.
Article in Malayalam | WPRIM | ID: wpr-629998

ABSTRACT

Chondrocytes were isolated from normal and microtic human auricular cartilage after ear surgery carried out at Universiti Kebangsaan Malaysia Medical Centre. Chondrocytes were cultured and expanded until passage 4. After reached confluence, cultured chondrocytes at each passage (P1, P2, P3 and P4) were harvested and assigned for growth profile analysis. There was no significant difference in cell viability between both normal and microtic samples (p = 0.84). Both samples showed no significant differences for growth profile parameters in terms of growth rate, population doubling time and total number of cell doubling, except in passage 1, where there is significant difference in cell growth rate (p = 0.004). This preliminary data has indicated that chondrocytes from microtic cartilage has the potential to be used in the reconstruction of human pinna in the future.

4.
The Medical Journal of Malaysia ; : 115-116, 2008.
Article in Malayalam | WPRIM | ID: wpr-629997

ABSTRACT

A potential cure for hearing loss would be to regenerate hair cells by stimulating cells of the damaged inner ear sensory epithelia to proliferate and differentiate into hair cells. Here, we investigated the possibility to isolate, culture-expand and characterize the cells from the cochlea membrane of adult mice. Our results showed that the cultured cells isolated from mouse cochlea membrane were heterogenous in nature. Morphologically there were epithelial like cells, hair cell like, nerve cell like and fibroblastic cells observed in the culture. The cultured cells were immunopositive for specific hair cell markers including Myosin 7a, Calretinin and Espin.

5.
The Medical Journal of Malaysia ; : 113-114, 2008.
Article in Malayalam | WPRIM | ID: wpr-629996

ABSTRACT

Spinal cord, sciatic nerve, olfactory ensheathing cell and bone marrow derived mesenchymal stem cells were evaluated as an alternative source for tissue engineering of nerve conduit. All cell sources were cultured in alpha-MEM medium. Olfactory Ensheathing Cell (OEC) showed the best result with higher growth kinetic compared to the others. Spinal cord and sciatic nerve were positive for GFAP, OEC were positive for GFAP, S100b and anti-cytokeratin 18 but negative for anti-Human Fibroblast.

6.
The Medical Journal of Malaysia ; : 111-112, 2008.
Article in Malayalam | WPRIM | ID: wpr-629995

ABSTRACT

This study was conducted to explore the feasibility of culturing conjunctiva epithelial cells in serum-free and feeder layer-free culture system with regard to the cell morphology and immunocytochemistry of the rabbit bulbar, fornix and palpebral conjunctiva epithelia. The results showed that epithelium cells from all the three conjunctiva regions can be cultured in a serum-free and feeder layer-free environment. We obtained highest epithelial growth from fornix region with minimum invasion of fibroblast cells compared to other area. All cultured cells were stained positive for cytokeratin 19 and MUC5AC and negative for cytokeratin 3. These findings suggested that fornix was a better source of cells for the development of tissue engineered conjunctiva for future clinical application.

7.
The Medical Journal of Malaysia ; : 109-110, 2008.
Article in Malayalam | WPRIM | ID: wpr-629994

ABSTRACT

The present work was to determine the development and re-epithelization of bilayered corneal construct (BCC) in vitro and in vivo using scanning electron microscopy (SEM). The in vitro BCC was transplanted to the rabbit's eye and after 90 days the BCC was harvested and analyzed. The corneas were processed for morphology studies. The result indicates that the BICC that was transplanted for 90 days showed good development and re-epithelization of epithelial layer similar to the normal cornea.

8.
The Medical Journal of Malaysia ; : 47-48, 2008.
Article in Malayalam | WPRIM | ID: wpr-629974

ABSTRACT

The emergence of tissue engineering and stem cell research has created a tremendous response amongst scientist in Malaysia. However, despite the enthusiastic to embark on the research we have to carefully divert the research towards our needs. This is due to our responsibility to address the mounting problem of communicable diseases here and a very limited funding. As commercialization is a key objective the combination of products towards treating or diagnosing communicable and non-communicable diseases in the developing country is another important factor. The discussion here is mainly on the evolution of tissue engineering in Malaysia and taking a model of tissue engineering in otolaryngology.

9.
The Medical Journal of Malaysia ; : 39-40, 2008.
Article in Malayalam | WPRIM | ID: wpr-629972

ABSTRACT

Bone marrow derived Mesenchymal stem cells (MSCs) were evaluated as an alternative source for tissue engineering of peripheral nerves. Human MSCs were subjected to a series of treatment with a reducing agent, retinoic acid and a combination of trophic factors. This treated MSCs differentiated into Schwann cells were characterized in vitro via flow cytometry analysis and immunocytochemically. In contrast to untreated MSCs, differentiated MSCs expressed Schwann cell markers in vitro, as we confirmed by flow cytometry analysis and immunocytochemically. These results suggest that human MSCs can be induced to be a substitute for Schwann cells that may be applied for nerve regeneration since it is difficult to grow Schwann cells in vitro.

10.
The Medical Journal of Malaysia ; : 34-34, 2008.
Article in Malayalam | WPRIM | ID: wpr-629971

ABSTRACT

Normal tracheal mucociliary clearance is the key to maintaining the health and defense of respiratory airway. Therefore the present of cilia and mucous blanket are important for tracheal epithelium to function effectively. In the present study, we prepared a tissue engineered respiratory epithelium construct (TEREC) made of autologous respiratory epithelium cells, fibroblast and fibrin from sheep owns blood which replaced a created tracheal mucosal defect. Scanning electron microscopy (SEM) showed encouraging result where immature cilia were present on the surface of TEREC. This result indicates that engineered respiratory epithelium was able to function as normal tissue.

11.
The Medical Journal of Malaysia ; : 32-33, 2008.
Article in Malayalam | WPRIM | ID: wpr-629970

ABSTRACT

The angiogenic potential of native skin (NS), keratinocytes single skin equivalent (SSE-K), fibroblasts single skin equivalent (SSE-F) and bilayered skin equivalent secreting angiogenic growth factors such as transforming growth factor beta1 (TGF-beta1), vascular endothelial growth factor (VEGF), keratinocyte growth factor (KGF) and basic fibroblast growth factor (bFGF) in the in vitro systems at 24, 48, 72 hours and 7 days was compared using Enzyme-Linked Immunosorbent Assay (ELISA). Bilayered skin equivalent exhibit highest release of growth factors within 24 hours to 7 days of culture compared to NS, SSE-K and SSE-F. This proved the potential of bilayered skin equivalent in producing and sustaining growth factors release to enhance angiogenesis, fibroblasts proliferation, matrix deposition, migration and growth of keratinocytes.

12.
The Medical Journal of Malaysia ; : 30-31, 2008.
Article in Malayalam | WPRIM | ID: wpr-629966

ABSTRACT

A major factor limiting survival following extensive thermal injury is insufficient availability of donor sites to provide enough skin for the required grafting procedures. Limitation of autologous grafting promotes the usage of allograft skin substitutes to promote wound healing. Here, we investigated the wound healing potential of allograft single layered tissue engineered skin which comprises of either keratinocytes (SLTES-K) or fibroblast (SLTES-F) with fibrin as the delivery system. Results from gross and microscopic evaluation showed our single layered tissue engineered skin constructed with keratinocytes or fibroblast after gamma radiation with the dosage of 2Gy could serve as allograft for the treatment of skin loss.

13.
The Medical Journal of Malaysia ; : 7-8, 2008.
Article in Malayalam | WPRIM | ID: wpr-629961

ABSTRACT

Nerve stem cells have a unique characteristic in that they form spherical aggregates, also termed neurospheres, in vitro. The study demonstrated the successful derivation of these neurospheres from bone marrow culture. Their plasticity as nerve stem cells was confirmed. The findings further strengthens the pluripotency of cell populations within the bone marrow.

14.
The Medical Journal of Malaysia ; : 27-28, 2008.
Article in Malayalam | WPRIM | ID: wpr-629954

ABSTRACT

Tissue engineering applies the principle of engineering and life sciences towards the development of biological substitute that restore, maintain or improve tissue or organ function. Scientists grow tissues or organs in vitro and implant them when the body is unable to prompt into healing itself. This presentation aims to highlight the potential clinical application of engineered tissues being researched on at the Tissue Engineering Centre, Universiti Kebangsaan Malaysia Medical Centre.

15.
The Medical Journal of Malaysia ; : 9-10, 2008.
Article in Malayalam | WPRIM | ID: wpr-629914

ABSTRACT

Human adipose-derived stem cells (HADSC) have demonstrated the capacity of differentiating into bone depending on the specific induction stimuli and growth factors. However, investigation on stem cell characteristic after osteogenic differentiation is still lacking. The goal of this study was to investigate the differential expression of sternness and osteogenic genes in non-induced HADSC compared with HADSC after osteogenic induction using quantitative Real Time RT-PCR. Our results showed that OCT-4, REX-1, FZD9, OSC, RUNX, and ALP were up regulated after osteogenic induction. This may indicated that HADSCs after osteogenic induction still possessed some stemness properties.

16.
The Medical Journal of Malaysia ; : 196-197, 2004.
Article in Malayalam | WPRIM | ID: wpr-629962

ABSTRACT

Bone marrow harvested by aspiration contains connective tissue progenitor cells which can be selectively isolated and induced to express bone phenotype in vitro. The osteoblastic progenitor can be estimated by counting the number of cells attach using the haemacytometer. This study was undertaken to test the hypothesis that human aging is associated with a significant change on the number of osteoblastic progenitors in the bone marrow. Bone marrow aspirates were harvested from 38 patients, 14 men (age 11-70) and 24 women (age 10-70) and cultured in F12: DMEM (1:1). In total 15 bone marrow samples have been isolated from patients above 40 years old (men/women) of age. Fourteen (93.3%) of this samples failed to proliferate. Only one (6.7%) bone marrow sample from a male patient, aged 59 years old was successfully cultured. Seventy percent (16/23) of the samples from patient below than 40 years old were successfully cultured. However, our observation on the survival rate for cells of different gender from patient below 40 years old does not indicate any significant difference. From this study, we conclude that the growth of bone marrow stromal cells possibly for bone engineering is better from bone marrow aspirates of younger patient.


Subject(s)
Age Factors , Bone Marrow Cells/cytology , Cellular Senescence/physiology , Cell Division/physiology , Cell Survival/physiology , Mesenchymal Stem Cells/cytology , Osteoblasts/cytology , Sex Factors , Stromal Cells/cytology , Tissue Engineering
17.
The Medical Journal of Malaysia ; : 190-191, 2004.
Article in Malayalam | WPRIM | ID: wpr-629960

ABSTRACT

This study was to assess collagen type II and collagen type I gene expression in tissue-engineered human auricular: cartilage formed via tissue engineering technique. Large-scale culture expansions were transformed into 3D in vitro construct and were implanted subcutaneously on the dorsal of athymic mice. After 8 weeks, explanted construct was processed in the same manner of native cartilage to facilitate cells for gene expression analysis. Isolated cells from in vivo construct demonstrated expression of type II collagen gene comparable to native cartilage. This study verified that tissue-engineered auricular cartilage expressed cartilage specific gene, collagen type II after in vivo maturation.


Subject(s)
Actins/genetics , Cartilage/transplantation , Cellular Senescence/physiology , Cells, Cultured , Chondrocytes/cytology , Collagen Type I/genetics , Collagen Type II/genetics , Ear, External , Fibroblasts/cytology , Gene Expression/physiology , Mice, Nude , Phenotype , Reverse Transcriptase Polymerase Chain Reaction , Tissue Engineering/methods
18.
The Medical Journal of Malaysia ; : 188-189, 2004.
Article in Malayalam | WPRIM | ID: wpr-629959

ABSTRACT

Cartilage is regularly needed for reconstructive surgery. Basic research in tissue engineering is necessary to develop its full potential. We presented here the expression profile of type II collagen gene and type I collagen gene in human auricular monolayer culture expansion. Cultured chondrocytes documented a reduction in the expression level of collagen type II gene whilst collagen type I gene was gradually expressed through all the passages. This study demonstrated that human auricular chondrocytes lose its phenotypic expression during monolayer culture expansion. Further studies are required to enhance cartilage specific gene expression, collagen type II throughout the in vitro culture.


Subject(s)
Cells, Cultured , Chondrocytes/cytology , Collagen Type I/genetics , Collagen Type II/genetics , Ear, External , Fibroblasts/cytology , Phenotype , Reverse Transcriptase Polymerase Chain Reaction , Tissue Engineering/methods
19.
The Medical Journal of Malaysia ; : 186-187, 2004.
Article in Malayalam | WPRIM | ID: wpr-629958

ABSTRACT

Chitosan has similar structure to glycosaminoglycans in the tissue, thus may be a good candidates as tissue engineering scaffold. However, to improve their cell attachment ability, we try to incorporate this natural polymer with collagen by combining it via cross-linking process. In this preliminary study we evaluate the cell attachment ability of chitosan-collagen scaffold versus chitosan scaffold alone. Chitosan and collagen were dissolved in 1% acetic acid and then were frozen for 24 hours before the lyophilizing process. Human skin fibroblasts were seeded into both scaffold and were cultured in F12: DMEM (1:1). Metabolic activity assay were used to evaluate cell attachment ability of scaffold for a period of 1, 3, 7 and 14 days. Scanning electron micrographs shows good cell morphology on chitosan-collagen hybrid scaffold. In conclusion, the incorporation of collagen to chitosan will enhance its cell attachment ability and will be a potential scaffold in tissue engineering.


Subject(s)
Cell Adhesion/physiology , Chitosan , Collagen , Energy Metabolism/physiology , Fibroblasts/cytology , Microscopy, Electron, Scanning , Organ Culture Techniques/methods , Polymers , Tissue Engineering/methods
20.
The Medical Journal of Malaysia ; : 184-185, 2004.
Article in Malayalam | WPRIM | ID: wpr-629957

ABSTRACT

Scanning Electron Microscopy (SEM) and Transmission Electron Microscopy (TEM) evaluation were carried out in the in vivo skin construct using fibrin as biomaterial. To investigate its progressive remodeling, nude mice were grafted and the Extracellular Matrix (ECM) components were studied at four and eight weeks post-grafting. It was discovered that by 4 weeks of remodeling the skin construct acquired its native structure.


Subject(s)
Collagen/physiology , Extracellular Matrix/pathology , Fibroblasts/pathology , Mice, Nude , Microscopy, Electron, Scanning , Microscopy, Electron, Transmission , Regeneration/physiology , Skin/pathology , Skin Transplantation/pathology , Tissue Engineering
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