Displaying publications 21 - 40 of 277 in total

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  1. Insulin-transferrin-selenium prevent human chondrocyte dedifferentiation and promote the formation of high quality tissue engineered human hyaline cartilage
    Chua KH, Aminuddin BS, Fuzina NH, Ruszymah BH
    Eur Cell Mater, 2005 Jun 17;9:58-67; discussion 67.
    PMID: 15962238
    This study was to investigate the effects of insulin-transferrin-selenium (ITS) on the proliferation and quantitative gene expression of adult human nasal septum chondrocytes in monolayer culture expansion and the formation of tissue engineered hyaline cartilage. Effects of ITS on human nasal septum chondrocytes monolayer culture expansion and gene expression were evaluated in various culture media either added with 2% fetal bovine serum (FBS) or 1 ng/mL basic fibroblast growth factor plus 1 ng/mL transforming growth factor or both serum and growth factors supplementation in comparison with medium added with 10%FBS. Chondrocytes cultured in medium added with 2% fetal bovine serum and growth factors either supplemented with or without ITS were then mixed with pluronic F-127 hydrogel for in vivo tissue engineered cartilage formation in nude mice model. Engineered tissues were removed after 8 weeks of implantation and evaluated with histological staining, immunohistochemistry, transmission electron microscopy and quantitative gene expression analysis. ITS promoted human chondrocytes proliferation and reduced chondrocytes dedifferentiation in media supplemented with serum and growth factors. ITS with 2% FBS and growth factors provided 15-fold increased in chondrocytes number by the end of the culture period compared to the standard culture medium used in chondrocytes culture (medium added with 10% FBS). Engineered tissue resulted from ITS supplementation demonstrated higher quality of cartilage formation. In conclusion, our study has demonstrated the benefits of ITS supplementation in human chondrocytes monolayer culture and tissue engineering cartilage formation.
    Matched MeSH terms: Collagen Type I/genetics; Collagen Type II/genetics
  2. Formation of in vivo tissue engineered human hyaline cartilage in the shape of a trachea with internal support
    Ruszymah BH, Chua K, Latif MA, Hussein FN, Saim AB
    Int J Pediatr Otorhinolaryngol, 2005 Nov;69(11):1489-95.
    PMID: 15941595
    Treatment and management of congenital as well as post-traumatic trachea stenosis remains a challenge in pediatric surgery. The aim of this study was to reconstruct a trachea with human nasal septum chondrocytes by using the combination of biodegradable hydrogel and non-biodegradable high-density polyethylene (HDP) as the internal predetermined shape scaffold.
    Matched MeSH terms: Collagen Type II/metabolism
  3. Wound healing activity of Ocimum sanctum Linn with supportive role of antioxidant enzymes
    Shetty S, Udupa S, Udupa L, Somayaji N
    Indian J. Physiol. Pharmacol., 2006 Apr-Jun;50(2):163-8.
    PMID: 17051736
    The present study was performed to evaluate the wound healing and antioxidant effect of aqueous extract of Ocimum sanctum Linn. (O. sanctum) in rats. Albino rats of either sex were divided into 2 groups. Group I: Wounded control rats; Group II: Wounded rats administered O. sanctum aqueous extract. Wound breaking strength in incision wound model, epithelization period and percent wound contraction in excision wound model were studied. Using dead space wound model, granulation tissue breaking strength, granulation tissue dry weight, hydoxyproline level in dry granulation tissue, superoxide dismutase (SOD) and catalase levels in wet granulation tissue were estimated in both the groups. Increased wound breaking strength, decreased epithelization period, increased percent wound contraction, increased granulation tissue weight and hydroxyproline concentrations were observed. The increased activity of antioxidant enzymes such as SOD, catalase level in extract treated group compared to controls. Granulation tissue was subjected to histopathological examination to determine the pattern of lay-down for collagen using Haematoxylin and Eosin stains which confirm the results. Owing to wound healing and antioxidant activities, O. sanctum may be useful in the management of abnormal healing such as keloids and hypertrophic scars.
    Matched MeSH terms: Collagen/metabolism
  4. Fulltext An overview of bone cells and their regulating factors of differentiation
    Mohamed AM
    Malays J Med Sci, 2008 Jan;15(1):4-12.
    PMID: 22589609 MyJurnal
    Bone is a specialised connective tissue and together with cartilage forms the strong and rigid endoskeleton. These tissues serve three main functions: scaffold for muscle attachment for locomotion, protection for vital organs and soft tissues and reservoir of ions for the entire organism especially calcium and phosphate. One of the most unique and important properties of bone is its ability to constantly undergo remodelling even after growth and modelling of the skeleton have been completed. Remodelling processes enable the bone to respond and adapt to changing functional situations. Bone is composed of various types of cells and collagenous extracellular organic matrix, which is predominantly type I collagen (85-95%) called osteoid that becomes mineralised by the deposition of calcium hydroxyapatite. The non-collagenous constituents are composed of proteins and proteoglycans, which are specific to bone and the dental hard connective tissues. Maintenance of appropriate bone mass depends upon the precise balance of bone formation and bone resorption which is facilitated by the ability of osteoblastic cells to regulate the rate of both differentiation and activity of osteoclasts as well as to form new bone. An overview of genetics and molecular mechanisms that involved in the differentiation of osteoblast and osteoclast is discussed.
    Matched MeSH terms: Collagen Type I
  5. The use of fibrin and poly(lactic-co-glycolic acid) hybrid scaffold for articular cartilage tissue engineering: an in vivo analysis
    Munirah S, Kim SH, Ruszymah BH, Khang G
    Eur Cell Mater, 2008 Feb 21;15:41-52.
    PMID: 18288632
    Our preliminary results indicated that fibrin and poly(lactic-co-glycolic acid) (PLGA) hybrid scaffold promoted early chondrogenesis of articular cartilage constructs in vitro. The aim of this study was to evaluate in vivo cartilaginous tissue formation by chondrocyte-seeded fibrin/PLGA hybrid scaffolds. PLGA scaffolds were soaked carefully, in chondrocyte-fibrin suspension, and polymerized by dropping thrombin-calcium chloride (CaCl2) solution. PLGA-seeded chondrocytes were used as a control. Resulting constructs were implanted subcutaneously, at the dorsum of nude mice, for 4 weeks. Macroscopic observation, histological evaluation, gene expression and sulphated-glycosaminoglycan (sGAG) analyses were performed at each time point of 1, 2 and 4 weeks post-implantation. Cartilaginous tissue formation in fibrin/PLGA hybrid construct was confirmed by the presence of lacunae and cartilage-isolated cells embedded within basophilic ground substance. Presence of proteoglycan and glycosaminoglycan (GAG) in fibrin/PLGA hybrid constructs was confirmed by positive Safranin O and Alcian Blue staining. Collagen type II exhibited intense immunopositivity at the pericellular matrices. Chondrogenic properties were further demonstrated by the expression of gene encoded cartilage-specific markers, collagen type II and aggrecan core protein. The sGAG production in fibrin/PLGA hybrid constructs was higher than in the PLGA group. In conclusion, fibrin/PLGA hybrid scaffold promotes cartilaginous tissue formation in vivo and may serve as a potential cell delivery vehicle and a structural basis for articular cartilage tissue-engineering.
    Matched MeSH terms: Collagen Type II/genetics; Collagen Type II/metabolism
  6. Fulltext Isolation, identification and quantification of differentially expressed proteins from cancerous and normal breast tissues
    Othman MI, Majid MI, Singh M, Man CN, Lay-Harn G
    Ann. Clin. Biochem., 2008 May;45(Pt 3):299-306.
    PMID: 18482919 DOI: 10.1258/acb.2007.007104
    Infiltrating ductal carcinoma (IDCA) is the most common type of breast cancer accounting for 85% of all invasive breast cancers.
    Matched MeSH terms: Collagen Type I/analysis; Collagen Type I/isolation & purification
  7. In-vitro expression of adhesion molecules and bone specific markers are depending on the cell culture material
    Salin N, Ishak AK, Abdul Rahman S, Ali M, Nawawi HM, Said MS, et al.
    Med J Malaysia, 2008 Jul;63 Suppl A:67-8.
    PMID: 19024987
    Bone formation is an active process whereby osteoblasts are found on the surface of the newly formed bone. Adhesion to extracellular matrix is essential for the development of bone however not all surfaces are suitable for osteoblast adhesion and don't support osteoblastic functions. The objective of this study was to test the suitability of a collagen based microcarrier which would support osteoblastic functions.
    Matched MeSH terms: Collagen*
  8. Use of dermal regeneration template (INTEGRA) in reconstructive burn surgery
    Sasidaran R, Dorai AA, Sulaiman WA, Halim AS
    Med J Malaysia, 2008 Jul;63 Suppl A:29.
    PMID: 19024967
    We present our two year experience with a dermal regeneration template (INTEGRA) in burn reconstructive surgery for contracture release as well as a reconstructive tool for management of soft tissue loss.
    Matched MeSH terms: Collagen*
  9. Cultured epidermal autografts in combination with MEEK Micrografting technique in the treatment of major burn injuries
    Dorai AA, Lim CK, Fareha AC, Halim AS
    Med J Malaysia, 2008 Jul;63 Suppl A:44.
    PMID: 19024976
    The treatment of major burn injuries are a formidable challenge to the burn surgeon. Early aggressive surgery for deep to full thickness burn injuries is vital in the prevention of infection. The ultimate goal in major burn injuries is to prevent the onset of multi-resistant organisms and achieve early wound cover. The field of tissue engineering can help to expedite the healing of these burn wounds. The development of keratinocyte culture delivery system can be used clinically to fasten the healing process and save many lives.
    Matched MeSH terms: Collagen
  10. The effect of TGF-beta1 and beta-estradiol on glycosaminoglycan and type II collagen distribution in articular chondrocyte cultures
    Ab-Rahim S, Selvaratnam L, Kamarul T
    Cell Biol Int, 2008 Jul;32(7):841-7.
    PMID: 18479947 DOI: 10.1016/j.cellbi.2008.03.016
    Articular cartilage extracellular matrix (ECM) plays a crucial role in regulating chondrocyte functions via cell-matrix interaction, cytoskeletal organization and integrin-mediated signaling. Factors such as interleukins, basic fibroblast growth factor (bFGF), bone morphogenic proteins (BMPs) and insulin-like growth factor (IGF) have been shown to modulate the synthesis of extracellular matrix in vitro. However, the effects of TGF-beta1 and beta-estradiol in ECM regulation require further investigation, although there have been suggestions that these factors do play a positive role. To establish the role of these factors on chondrocytes derived from articular joints, a study was conducted to investigate the effects of TGF-beta1 and beta-estradiol on glycosaminoglycan secretion and type II collagen distribution (two major component of cartilage ECM in vivo). Thus, chondrocyte cultures initiated from rabbit articular cartilage were treated with 10ng/ml of TGF-beta1, 10nM of beta-estradiol or with a combination of both factors. Sulphated glycosaminoglycan (GAG) and type II collagen levels were then measured in both these culture systems. The results revealed that the synthesis of GAG and type II collagen was shown to be enhanced in the TGF-beta1 treated cultures. This increase was also noted when TGF-beta1 and beta-estradiol were both used as culture supplements. However, beta-estradiol alone did not appear to affect GAG or type II collagen deposition. There was also no difference between the amount of collagen type II and GAG being expressed when chondrocyte cultures were treated with TGF-beta1 when compared with cultures treated with combined factors. From this, we conclude that although TGF-beta1 appears to stimulate chondrocyte ECM synthesis, beta-estradiol fails to produce similar effects. The findings of this study confirm that contrary to previous claims, beta-estradiol has little or no effect on chondrocyte ECM synthesis. Furthermore, the use of TGF-beta1 may be useful in future studies looking into biological mechanisms by which ECM synthesis in chondrocyte cultures can be augmented, particularly for clinical application.
    Matched MeSH terms: Collagen Type II/metabolism*
  11. Fibrin promotes proliferation and matrix production of intervertebral disc cells cultured in three-dimensional poly(lactic-co-glycolic acid) scaffold
    Sha'ban M, Yoon SJ, Ko YK, Ha HJ, Kim SH, So JW, et al.
    J Biomater Sci Polym Ed, 2008;19(9):1219-37.
    PMID: 18727862 DOI: 10.1163/156856208785540163
    Previously, we have proven that fibrin and poly(lactic-co-glycolic acid) (PLGA) scaffolds facilitate cell proliferation, matrix production and early chondrogenesis of rabbit articular chondrocytes in in vitro and in vivo experiments. In this study, we evaluated the potential of fibrin/PLGA scaffold for intervertebral disc (IVD) tissue engineering using annulus fibrosus (AF) and nucleus pulposus (NP) cells in relation to potential clinical application. PLGA scaffolds were soaked in cells-fibrin suspension and polymerized by dropping thrombin-sodium chloride (CaCl(2)) solution. A PLGA-cell complex without fibrin was used as control. Higher cellular proliferation activity was observed in fibrin/PLGA-seeded AF and NP cells at each time point of 3, 7, 14 and 7 days using the MTT assay. After 3 weeks in vitro incubation, fibrin/PLGA exhibited a firmer gross morphology than PLGA groups. A significant cartilaginous tissue formation was observed in fibrin/PLGA, as proven by the development of cells cluster of various sizes and three-dimensional (3D) cartilaginous histoarchitecture and the presence of proteoglycan-rich matrix and glycosaminoglycan (GAG). The sGAG production measured by 1,9-dimethylmethylene blue (DMMB) assay revealed greater sGAG production in fibrin/PLGA than PLGA group. Immunohistochemical analyses showed expressions of collagen type II, aggrecan core protein and collagen type I genes throughout in vitro culture in both fibrin/PLGA and PLGA. In conclusion, fibrin promotes cell proliferation, stable in vitro tissue morphology, superior cartilaginous tissue formation and sGAG production of AF and NP cells cultured in PLGA scaffold. The 3D porous PLGA scaffold-cell complexes using fibrin can provide a vehicle for delivery of cells to regenerate tissue-engineered IVD tissue.
    Matched MeSH terms: Collagen Type I/genetics; Collagen Type I/metabolism; Collagen Type II/genetics; Collagen Type II/metabolism
  12. Fulltext Desmoplastic Ameloblastoma of the Mandible: A Case Report
    Rajan Saini, Abdul Rani Samsudin
    International Medical Journal Malaysia, 2008;7(1):1-7.
    MyJurnal
    Desmoplastic ameloblastoma (DA) is a relatively rare histological variant of ameloblastoma. DA do not present with radiographic or clinical features that are typical of other variants of ameloblastoma. On gross examination, DA appears as a solid mass, unlike the conventional ameloblastoma that contains fluid-filled spaces. Although radiographic examination of ameloblastomas usually reveals unilocular or multilocular radiolucency, DA may appear as a mixed radiopaque-radiolucent lesion. Histologically, DA is characterized by small nests and strands of “compressed” odontogenic epithelium supported by pronounced collagenized stroma. This report describes the case of a 30-year-old male with DA of the left mandible.
    Matched MeSH terms: Collagen
  13. The microscopic biological response of human chondrocytes to bovine bone scaffold
    Abdullah B, Shibghatullah AH, Hamid SS, Omar NS, Samsuddin AR
    Cell Tissue Bank, 2009 Aug;10(3):205-13.
    PMID: 18975136 DOI: 10.1007/s10561-008-9111-2
    This study was performed to determine the microscopic biological response of human nasal septum chondrocytes and human knee articular chondrocytes placed on a demineralized bovine bone scaffold. Both chondrocytes were cultured and seeded onto the bovine bone scaffold with seeding density of 1 x 105 cells per 100 microl/scaffold and incubated for 1, 2, 5 and 7 days. Proliferation and viability of the cells were measured by mitochondrial dehydrogenase activity (MTT assay), adhesion study was analyzed by scanning electron microscopy and differentiation study was analyzed by immunofluorescence staining and confocal laser scanning electron microscopy. The results showed good proliferation and viability of both chondrocytes on the scaffolds from day 1 to day 7. Both chondrocytes increased in number with time and readily grew on the surface and into the open pores of the scaffold. Immunofluorescence staining demonstrated collagen type II on the scaffolds for both chondrocytes. The results showed good cells proliferation, attachment and maturity of the chondrocytes on the demineralized bovine bone scaffold. The bovine bone being easily resourced, relatively inexpensive and non toxic has good potential for use as a three dimensional construct in cartilage tissue engineering.
    Matched MeSH terms: Collagen Type II/biosynthesis
  14. Fulltext Common laboratory tests for rheumatological disorders: how do they help the diagnosis?
    Das Gupta E
    Malays Fam Physician, 2009;4(2-3):48-50.
    PMID: 25606162 MyJurnal
    No screening test is ideal for detecting rheumatic diseases; diagnosis depends on appropriate history and thorough physical examination. Sometimes, laboratory investigations may be useful in confirming or ruling out rheumatic disease after a clinical diagnosis is considered. Once a rheumatic disease has been diagnosed, certain laboratory tests can help in assessing prognosis or determining the extent of the disease. Laboratory tests may also help the physician monitor certain rheumatic diseases, guide treatment or assess potential drug toxicity.
    Matched MeSH terms: Collagen Diseases
  15. Fulltext Histological observation of gingival enlargement induced by cyclosporin a and nifedipine: a study in rabbits
    Al-Bayaty, F.H., Al-Tay, B.O., Al-Kushali, S.S., Mahmmod, L.
    ASM Science Journal, 2009;3(1):45-50.
    MyJurnal
    A study was undertaken to estimate the histological changes of gingival enlargement induced by
    Cyclosporin A (CsA) and Nifedipine, separately and in combination. Twelve adult rabbits were divided equally into four main groups. The first group received 10 mg/kg/day Nifedipine, the second received 10 mg/kg/day CsA, and the third received a combination of 10 mg/kg /day Nifedipine and CsA by gastric feeding. The fourth was regarded as a control group. Animals were given the drugs from day 1 of the experiment until day 70. They were then sacrificed for histological purposes. Results showed increase in the thickness of the epithelium with keratosis and acanthosis, and also increased vascularity. Collagen fibres and fibroblasts at different rates in the three histological groups were observed. Significant alveolar bone resorption with increased marrow spaces filled with fatty tissue were found in the CsA group. Non-significant changes in the alveolar bone of the Nifedipine group while subsequent bone resorption and bone deposition were seen in the combination group. These changes could be due to the effect of both drugs. Significant changes in the gingiva and the alveolar bone were shown in the three experimental groups compared with the control group.
    Matched MeSH terms: Collagen
  16. Fulltext Acceleration of wound healing by orthosiphon stamineus leaf extract in rats
    Mahmood, A.A., Hapipah, M.A., Noor, S.M., Kuppusamy, U.R., Salmah, I., Salmah, I., et al.
    ASM Science Journal, 2009;3(1):51-57.
    MyJurnal
    The effects of topical application of Orthosiphon stamineus leaf extract on the rate of wound healing and histology of the healed wound were assessed. Four groups of adult male Sprague Dawley rats were experimentally wounded in the posterior neck area. A thin layer of blank placebo was applied topically to wounds of Group 1 rats. Wounds of experimental animals (Group 2 and 3) were dressed with placebo containing 5% and 10% O. stamineus extract, respectively. A thin layer of Intrasite gel® was applied topically to wounds of Group 4 animals as reference. Macroscopically, wounds dressed with placebo containing 5% (healed on day 14.50 ± 0.43) and 10% (healed on day 13.83 ± 0.21) O. stamineus extract each or Intrasite gel® (healed on day 13.13 ± 0.42) significantly accelerated the rate of wound healing compared to wounds dressed with blank placebo. Histological analysis of healed wounds confirmed the results. Wounds dressed with placebo containing 5%, 10% O.stamineus or Intrasite gel® showed markedly less scar width at wound enclosure and granulating tissue contained markedly more collagen, proliferating fibroblast with angiogenesis, and no inflammatory cells compared to wounds dressed with blank placebo. In conclusion, placebo containing 5% or 10% O. stamineus on extract-dressed wounds significantly accelerated the rate of wound healing in rats.
    Matched MeSH terms: Collagen
  17. Anti-arthritic potential of the plant Justicia gendarussa Burm F
    Paval J, Kaitheri SK, Potu BK, Govindan S, Kumar RS, Narayanan SN, et al.
    Clinics (Sao Paulo), 2009;64(4):357-62.
    PMID: 19488595
    OBJECTIVE: To evaluate the anti-arthritic potential of the plant Justicia gendarussa using two different rat models.

    MATERIALS AND METHOD: The anti-arthritic potential of the alcoholic extract of the plant Justicia gendarussa was evaluated using the Freund's adjuvant-induced and collagen-induced arthritic rat models. The rats were treated with the ethanolic extract of Justicia gendarussa and with standard aspirin.

    RESULTS: The ethanolic extract of Justicia gendarussa showed significant anti-arthritic activity that was statistically similar to that of aspirin. Our results suggest that the alcoholic extract of Justicia gendarussa exhibits significant anti-arthritic potential.

    Matched MeSH terms: Collagen Type II
  18. Tissue engineering approach to repair abdominal wall defects using cell-seeded bovine tunica vaginalis in a rabbit model
    Ayele T, Zuki AB, Noorjahan BM, Noordin MM
    J Mater Sci Mater Med, 2010 May;21(5):1721-30.
    PMID: 20135201 DOI: 10.1007/s10856-010-4007-7
    The aim of this study was to engineer skeletal muscle tissue for repair abdominal wall defects. Myoblast were seeded onto the scaffolds and cultivated in vitro for 5 days. Full thickness abdominal wall defects (3 x 4 cm) were created in 18 male New Zealand white rabbits and randomly divided into two equal groups. The defects of the first group were repaired with myoblast-seeded-bovine tunica vaginalis whereas the second group repaired with non-seeded-bovine tunica vaginalis and function as a control. Three animals were sacrificed at 7th, 14th, and 30th days of post-implantation from each group and the explanted specimens were subjected to macroscopic and microscopic analysis. In every case, seeded scaffolds have better deposition of newly formed collagen with neo-vascularisation than control group. Interestingly, multinucleated myotubes and myofibers were only detected in cell-seeded group. This study demonstrated that myoblast-seeded-bovine tunica vaginalis can be used as an effective scaffold to repair severe and large abdominal wall defects with regeneration of skeletal muscle tissue.
    Matched MeSH terms: Collagen
  19. Fulltext Urinary type IV collagen levels in diabetes mellitus
    Sthaneshwar P, Chan SP
    Malays J Pathol, 2010 Jun;32(1):43-7.
    PMID: 20614725 MyJurnal
    Type IV collagen is the principal component of glomerular basement membrane and messangial matrix. Studies have shown increased levels of urinary type IV collagen (uIV) in diabetic patients compared to healthy controls. The concentration of uIV increases gradually as diabetic nephropathy progresses.
    AIM AND METHOD: This study was carried out to determine whether urinary type IV collagen (uIV) can serve as an indicator of diabetic nephropathy. Using a sandwich enzyme immunoassay technique, uIV excretion was determined in 30 type 2 diabetic patients with normoalbuminuria and 20 patients with microalbuminuria.
    RESULTS: uIV excretion was significantly increased in type 2 diabetics, in both normoalbuminuric and microalbuminuric patients, compared with healthy controls. The increase in urinary type IV collagen was well correlated with the amount of urinary albumin but not with HbA1C.
    CONCLUSION: Our findings that uIV is higher in those with microalbuminuria and correlates with albuminuria, support uIV as an indicator of diabetic nephropathy. Whether the increased uIV excretion would predict the impending renal failure needs further confirmation.
    Matched MeSH terms: Collagen Type IV/urine*
  20. Expansion of human articular chondrocytes and formation of tissue-engineered cartilage: a step towards exploring a potential use of matrix-induced cell therapy
    Munirah S, Samsudin OC, Aminuddin BS, Ruszymah BH
    Tissue Cell, 2010 Oct;42(5):282-92.
    PMID: 20810142 DOI: 10.1016/j.tice.2010.07.002
    Monolayer culture expansion remains as a fundamental step to acquire sufficient number of cells for 3D constructs formation. It has been well-documented that cell expansion is however accompanied by cellular dedifferentiation. In order to promote cell growth and circumvent cellular dedifferentiation, we evaluated the effects of Transforming Growth Factor Beta-2 (TGF-β2), Insulin-like Growth Factor-I (IGF-I) and basic Fibroblast Growth Factor (bFGF) combination on articular chondrocytes culture and 'chondrocytes-fibrin' construct formation. Chondrocytes were serially cultured in: (1) F12:DMEM+10% Foetal Bovine Serum (FBS) with growth factors (FD10GFs), (2) F12:DMEM+2%FBS with the growth factors (FD2GFs) and, (3) F12:DMEM+10%FBS without growth factors (FD) as control. Cultured chondrocytes were evaluated by means of growth kinetics parameters, cell cycle analysis, quantitative phenotypic expression of collagen type II, aggrecan core protein sox-9 and collagen type I and, immunochemistry technique. Harvested chondrocytes were incorporated with plasma-derived fibrin and were polymerized to form the 3D constructs and implanted subcutaneously at the dorsum of athymic nude mice for eight (8) weeks. Resulted constructs were assigned for gross inspections and microscopic evaluation using standard histochemicals staining, immunochemistry technique and, quantitative phenotypic expression of cartilage markers to reassure cartilaginous tissue formation. Growth kinetics performance of chondrocytes cultured in three (3) types of culture media from the most to least was in the following order: FD10GFs>FD2GFs>FD. Following growth kinetics analysis, we decided to use FD10GFs and FD (control) for further evaluation and 'chondrocytes-fibrin' constructs formation. Chondrocytes cultured in FD10GFs preserved the normal diploid state (2c) with no evidence of aneuploidy, haploidy or tetraploidy. Expression of cartilage-specific markers namely collagen type II, aggrecan core protein and sox-9 were significantly higher in FD10GFs when compared to control. After implantation, 'chondrocytes-fibrin' constructs exhibited firm, white, smooth and glistening cartilage-like properties. FD10GFs constructs formed better quality cartilage-like tissue than FD constructs in term of overall cartilaginous tissue formation, cells organization and extracellular matrix distribution in the specimens. Cartilaginous tissue formation was confirmed by the presence of lacunae and cartilage-isolated cells embedded within basophilic ground substance. Presence of proteoglycan was confirmed by positive Safranin O staining. Collagen type II exhibited immunopositivity at the pericellular and inter-territorial matrix area. Chondrogenic properties of the construct were further confirmed by the expression of genes encoding collagen type II, aggrecan core protein and sox9. In conclusion, FD10GFs promotes the proliferation of chondrocytes and formation of good quality 'chondrocytes-fibrin' constructs which may have potential use of matrix-induced cell implantation.
    Matched MeSH terms: Collagen Type II/biosynthesis
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