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Bone marrow mesenchymal stem cells differentiation and proliferation on the surface of coral implant

Al-Salihi KA, Samsudin AR

Med J Malaysia, 2004 May;59 Suppl B:45-6.
PMID: 15468811

This study was designed to evaluate the ability of natural coral implant to provide an environment for marrow cells to differentiate into osteoblasts and function suitable for mineralized tissue formation. DNA content, alkaline phosptatase (ALP) activity, calcium (Ca) content and mineralized nodules, were measured at day 3, day 7 and day 14, in rat bone marrow stromal cells cultured with coral discs glass discs, while cells alone and coral disc alone were cultured as control. DNA content, ALP activity, Ca content measurements showed no difference between coral, glass and cells groups at 3 day which were higher than control (coral disc alone), but there were higher measurement at day 7 and 14 in the cell cultured on coral than on glass discs, control cells and control coral discs. Mineralized nodules formation (both in area and number) was more predominant on the coral surface than in control groups. These results showed that natural coral implant provided excellent and favorable situation for marrow cell to differentiate to osteoblasts, lead to large amount of mineralized tissue formation on coral surface. This in vitro result could explain the rapid bone bonding of coral in vivo.
Fulltext Pharmacological inhibition of endoplasmic reticulum stress mitigates osteoporosis in a mouse model of hindlimb suspension

Al-Daghestani H, Qaisar R, Al Kawas S, Ghani N, Rani KGA, Azeem M, et al.

Sci Rep, 2024 Feb 27;14(1):4719.
PMID: 38413677 DOI: 10.1038/s41598-024-54944-7

Hindlimb suspension (HLS) mice exhibit osteoporosis of the hindlimb bones and may be an excellent model to test pharmacological interventions. We investigated the effects of inhibiting endoplasmic reticulum (ER) stress with 4-phenyl butyrate (4-PBA) on the morphology, physicochemical properties, and bone turnover markers of hindlimbs in HLS mice. We randomly divided 21 male C57BL/6J mice into three groups, ground-based controls, untreated HLS group and 4-PBA treated group (HLS+4PBA) (100mg/kg/day, intraperitoneal) for 21 days. We investigated histopathology, micro-CT imaging, Raman spectroscopic analysis, and gene expression. Untreated HLS mice exhibited reduced osteocyte density, multinucleated osteoclast-like cells, adipocyte infiltration, and reduced trabecular striations on micro-CT than the control group. Raman spectroscopy revealed higher levels of ER stress, hydroxyproline, non-collagenous proteins, phenylalanine, tyrosine, and CH2Wag as well as a reduction in proteoglycans and adenine. Furthermore, bone alkaline phosphatase and osteocalcin were downregulated, while Cathepsin K, TRAP, and sclerostin were upregulated. Treatment with 4-PBA partially restored normal bone histology, increased collagen crosslinking, and mineralization, promoted anti-inflammatory markers, and downregulated bone resorption markers. Our findings suggest that mitigating ER stress with 4-PBA could be a therapeutic intervention to offset osteoporosis in conditions mimicking hindlimb suspension.
Three-Dimensional Radiological Assessment of Alveolar Bone Volume Preservation Using Bovine Bone Xenograft

Al Qabbani A, Al Kawas S, A Razak NH, Al Bayatti SW, Enezei HH, Samsudin AR, et al.

J Craniofac Surg, 2018 Mar;29(2):e203-e209.
PMID: 29303859 DOI: 10.1097/SCS.0000000000004263

INTRODUCTION: Alveolar bone is critical in supporting natural teeth, dental implants as well as a removable and fixed prosthesis. Alveolar bone volume diminishes when its associated natural tooth is lost.
OBJECTIVE: The aim of this study is to evaluate the effectiveness of bovine bone granules on alveolar bone socket augmentation for ridge preservation following atraumatic tooth extraction.
MATERIALS AND METHODS: Twenty medically fit patients (12 males and 8 females aged between 18 and 40 years) who needed noncomplicated tooth extraction of 1 mandibular premolar tooth were divided randomly and equally into 2 groups. In control group I, the empty extraction socket was left untreated and allowed to heal in a conventional way. In group II, the empty extraction socket wound was filled with lyophilized bovine bone xenograft granules 0.25 to 1 mm of size, 1 mL/vial. A resorbable pericardium membrane was placed to cover the defect. Clinical and 3-dimensional radiological assessments were performed at day 0, 3 months, and 9 months postoperative.
RESULTS: There were no clinical differences in general wound healing between the groups. Comparisons within the groups showed a significant difference of bone resorption of 1.49 mm (95% confidence interval, 0.63-2.35) at 3 months, and further resorption of 1.84 mm (P ≤ 0.05) at 9 months in the control group. No significant changes of bone resorption were observed in group II during the same time interval. Comparison between groups showed a significant difference of bone resorption at 3 and 9 months (2.40 and 2.88 mm, respectively).
CONCLUSION: The use of lyophilized demineralized bovine bone granules in socket preservation to fill in the extraction socket seems essential in preserving the alveolar bone dimension as it showed excellent soft and hard tissue healing. This study concludes that the alveolar bone socket exhibited a dynamic process of resorption from the first day of tooth extraction. Evidence shows the possibility of using bovine bone granules routinely in socket volume preservation techniques following tooth extraction.
Biomechanical and radiological assessment of immediate implants for alveolar ridge preservation

Al Qabbani A, Al Kawas S, Enezei H, Razak NHA, Al Bayatti SW, Samsudin AR, et al.

Dent Res J (Isfahan), 2018;15(6):420-429.
PMID: 30534170

BACKGROUND: The aim of this study was to evaluate the stability of immediate implant placement for alveolar bone augmentation and preservation with bovine bone graft following atraumatic tooth extraction.
MATERIALS AND METHODS: This was a prospective interventional study with convenient sampling (n = 10). Thirty patients aged between 18 and 40 years, who needed noncomplicated tooth extraction of mandibular premolar tooth, were sequentially divided equally into three groups. In Group I, simple extraction was done and the empty extraction socket left to heal conventionally. In Group II, extraction sockets were filled with lyophilized bovine granules only. In Group III, immediate implants were placed into extraction sockets, and the buccal gap was also filled with bovine granules. All groups were subjected to cone beam computed tomography scan for radiological evaluation. Assessment of biomechanical stability (radiofrequency analysis [RFA] was performed at 9 months postoperative for Group III to assess the degree of secondary stability of the implants using Osstell. Repeated measure analysis of variance (ANOVA) test was applied when comparing within each group at three different time intervals, whereas one-way ANOVA was applied followed by post hoc-tukey test when comparing between groups. P < 0.05 was considered statistically significant.
RESULTS: Radiological assessment reveals a significant difference of bone resorption in alveolar dimension within Group I; 1.49 mm (P = 0.002), and 0.82 mm (P = 0.005), respectively, between day 0 and 3 months. Comparison between Group I and III showed a highly significant difference of bone resorption in ridge width at 3 months 2.56 mm (P = 0.001) and at 9 months interval 3.2 mm (P < 0.001). High RFA values demonstrating an excellent biomechanical stability were observed in Group III at 9 months postoperatively.
CONCLUSION: The insertion of immediate implants in extraction sockets with bovine bone augmentation of the buccal gap was able to preserve a greater amount of alveolar ridge volume.
Fulltext Evaluation of decellularization process for developing osteogenic bovine cancellous bone scaffolds in-vitro

Al Qabbani A, Rani KGA, Syarif J, AlKawas S, Sheikh Abdul Hamid S, Samsudin AR, et al.

PLoS One, 2023;18(4):e0283922.
PMID: 37018321 DOI: 10.1371/journal.pone.0283922

Current immunological issues in bone grafting regarding the transfer of xenogeneic donor bone cells into the recipient are challenging the industry to produce safer acellular natural matrices for bone regeneration. The aim of this study was to investigate the efficacy of a novel decellularization technique for producing bovine cancellous bone scaffold and compare its physicochemical, mechanical, and biological characteristics with demineralized cancellous bone scaffold in an in-vitro study. Cancellous bone blocks were harvested from a bovine femoral head (18-24 months old) subjected to physical cleansing and chemical defatting, and further processed in two ways. Group I was subjected to demineralization, while Group II underwent decellularization through physical, chemical, and enzymatic treatments. Both were then freeze-dried, and gamma radiated, finally producing a demineralized bovine cancellous bone (DMB) scaffold and decellularized bovine cancellous bone (DCC) scaffold. Both DMB and DCC scaffolds were subjected to histological evaluation, scanning electron microscopy/energy-dispersive X-ray spectroscopy (SEM/EDS), fourier-transform infrared spectroscopy (FTIR), quantification of lipid, collagen, and residual nucleic acid content, and mechanical testing. The osteogenic potential was investigated through the recellularization of scaffolds with human osteoblast cell seeding and examined for cell attachment, proliferation, and mineralization by Alizarin staining and gene expression. DCC produced a complete acellular extracellular matrix (ECM) with the absence of nucleic acid content, wider pores with extensive interconnectivity and partially retaining collagen fibrils. DCC demonstrated a higher cell proliferation rate, upregulation of osteogenic differentiation markers, and substantial mineralized nodules production. Our findings suggest that the decellularization technique produced an acellular DCC scaffold with minimal damage to ECM and possesses osteogenic potential through the mechanisms of osteoconduction, osteoinduction, and osteogenesis in-vitro.
Fulltext Evaluation of the osteogenic potential of demineralized and decellularized bovine bone granules following implantation in rat calvaria critical-size defect model

Al Qabbani A, Rani KGA, AlKawas S, Sheikh Abdul Hamid S, Yap Abdullah A, Samsudin AR, et al.

PLoS One, 2023;18(12):e0294291.
PMID: 38127838 DOI: 10.1371/journal.pone.0294291

The aim of this study was to compare the ability of demineralized (DMB) and decellularized (DCC) bovine bone granules to support bone regeneration in rat calvaria critical-size defects. DMB and DCC were prepared using a previously published method. The granule size used ranged between 500 and 750 μm. A total of forty-eight Sprague-Dawley rats were divided into two groups (n = 24). A pair of 5 mm diameter defects were created on the calvaria of the rats in the right and left parietal bone in both groups. Group A animals received DMB granules and Group B received DCC granules in the right parietal defect side while the left parietal untreated defect acted as sham surgery for both groups. Four animals per group were euthanized in a CO2 chamber at day 7, 14 and 21 post-surgery and the calvaria implantation site biopsy harvested was subjected to osteogenic gene expression analysis. Another four animals per group were euthanized at days 15, 30 and 60 post surgery and the calvaria implantation site biopsy harvested was subjected to histological, immunohistochemistry, RAMAN spectroscopy and Micro-CT analysis at the mentioned time points. Statistical analysis was conducted using t-tests and ANOVA. Histomorphometry showed significantly higher new bone formation in the DCC sites (p<0.05) compared to DMB. Both DMB and DCC implantation sites showed distinct staining for osteocalcin and osteopontin proteins compared to their respective sham sites. By day 21 after implantation, DCC sites demonstrated significantly elevated mRNA levels of osteonectin (p<0.001), osteopontin (p<0.001), osteocalcin (p<0.0001), ALP (p<0.01), and BMP-2 (p<0.001) compared to DMB. However, VEGF expression showed no significant differences at this time point between the two groups. Micro-CT analysis also showed enhanced defect closure and higher bone density in DCC implanted sites while RAMAN spectra demonstrated increased abundance of collagen and bone minerals, especially, PO43- ions than DMB. In conclusion, both DMB and DCC granules demonstrated favorable osteogenic potential in critical-sized defects, with DCC exhibited superior osteoconductive, osteoinductive and osteogenesis properties.
Fixed presurgical orthopaedics for bilateral cleft lip and palate

Abu-Rub N, Samsudin AR, Abdullah AB, Abdullah N

Aust Orthod J, 2005 May;21(1):39-43.
PMID: 16433080

Presurgical orthopaedics has been employed since the 1950s as an adjunctive neonatal therapy for the correction of cleft lip and palate. It is accepted that presurgical orthopaedic plates facilitate lip repair and balanced orofacial growth.
An in vivo study of a locally-manufactured hydroxyapatite-based material as bone replacement material

Abdul Razak NH, Al-Salihi KA, Samsudin AR

Med J Malaysia, 2004 May;59 Suppl B:119-20.
PMID: 15468847

Defects were created in the mandible of a rabbit model whereby the right side was implanted with hydroxyapatite (HA) while the left side was left empty to act as control. Both the implant and control sites were evaluated clinically and histologically at 4,12,20,22 weeks. Decalcified sections were studied under confocal laser scanning microscope. No reactive cells were evident microscopically in all sections. There was bone ingrowth as early as 4 weeks when viewed by the topographic method. Enhancement of osteoconduction was evident by the presence of abundant capillaries, perivascular tissue and osteoprogenitor cells of the host. At 22 weeks, the implanted defect showed mature bone formation filling almost the whole field. This study demonstrated that the dense HA exhibits excellent biocompatibility as noted by the complete absence of reactive cells. It also promotes osteoconduction.
Fulltext Dental anomalies and facial profile abnormality of the non-syndromic cleft lip and palate children in Kelantan

Abd Rahman N, Abdullah N, Samsudin AR, Naing Mohd Ayub Sadiq L

Malays J Med Sci, 2004 Jul;11(2):41-51.
PMID: 22973126

This study was done to determine the prevalence of dental anomalies and facial profile abnormality and its association with the non-syndromic cleft lip and palate (CLP) as compared to the non-cleft children. A comparative cross sectional study was conducted where the case group consist of 98 non-syndromic CLP children-unilateral (UCLP) and bilateral (BCLP) who attended the Combined Clinic at Kota Bharu Dental Clinic (KBDC) while the comparison group comprised of 109 non-cleft children who attended the outpatient clinic at KBDC. Their ages were between 3 to 12 years old. Clinical oral and facial profile examinations were carried out to look for dental anomalies (morphology, number and alignment of teeth) and facial profile abnormality. The prevalence of anomalies in morphology of teeth in CLP (24.5%) and non-cleft (10.1%), number of teeth in CLP (44.9%) and non-cleft (7.3%), mal-alignment in CLP (79.6%) and non-cleft (27.5%) and facial profile abnormality in CLP (26.5%) and non-cleft (9.1 %). There was a significant association between CLP and anomalies in morphology, number, mal-alignment and abnormality in facial profile; (p < 0.05). Therefore, there was a high prevalence and risk of dental anomalies and facial profile abnormality in the CLP children compared to the non-cleft children.