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Re: Kozacioglu et al.: Anatomy of the dorsal nerve of the penis, clinical implications (Urology 2014;83:121-125)

Suhaimi FH, Abd Ghafar N, Das S

Urology, 2014 May;83(5):1207-8.
PMID: 24767534 DOI: 10.1016/j.urology.2014.01.033
Fulltext The effects of virgin coconut oil on fibroblasts and myofibroblasts on diabetic wound healing

Wong, S.K., Rangiah T., Bakri, N.S.A., Ismail, W.N.A, Bojeng, E.E.F., Abd Rahiman, M.A., et al.

Medicine & Health, 2019;14(2):132-141.
MyJurnal

Delayed wound healing is one of the major complications of diabetes mellitus where it often leads to amputation. Virgin coconut oil (VCO) is a natural oil widely used to treat wounds and burns traditionally. It possesses potent antioxidant and antibacterial activities. This study aimed to determine the effect of VCO on the fibroblast population in diabetic wound healing. Full thickness cutaneous wound tissues were collected from non-treated, VCO-treated, and silver sulfadiazine (SS)- treated diabetic rats. The tissues were then subjected to Verhoeff eosin staining and immunohistology of fibroblast and myofibroblast. Histological analysis showed increased collagen deposition with intact epidermis in the VCO treated group compared to decreased collagen deposition with damaged epidermis in both non-treated and SS-treated groups. Interestingly, more fibroblasts and myofibroblasts were observed in the non-treated group compared to the VCO- and SS-treated groups. VCO significantly promoted wound healing process in diabetic rats via promoting re-epithelialization, and increasing collagen fibres deposition and wound contraction. The results suggested VCO can be used to treat diabetic wounds.
Fulltext The role of BMP4 in adipose-derived stem cell differentiation: A minireview

Setiawan AM, Kamarudin TA, Abd Ghafar N

Front Cell Dev Biol, 2022;10:1045103.
PMID: 36340030 DOI: 10.3389/fcell.2022.1045103

Bone morphogenetic protein 4 (BMP4) is a member of the transforming growth factor beta (TGF-β) superfamily of cytokines responsible for stem cells' commitment to differentiation, proliferation, and maturation. To date, various studies have utilized BMP4 as a chemical inducer for in vitro differentiation of human mesenchymal stem cells (MSCs) based on its potential. BMP4 drives in vitro differentiation of ADSC via TGF-β signaling pathway by interactions with BMP receptors leading to the activation of smad-dependent and smad-independent pathways. The BMP4 signaling pathways are regulated by intracellular and extracellular BMP4 antagonists. Extracellular BMP4 antagonist prevents interaction between BMP4 ligand to its receptors, while intracellular BMP4 antagonist shutdowns the smad-dependent pathways through multiple mechanisms. BMP4 proved as one of the popular differentiation factors to induce ADSC differentiation into cell from mesodermal origin. However, addition of all-trans retinoic acid is also needed in trans-differentiation of ADSC into ectodermal lineage cells. Suggesting that both BMP4 and RA signaling pathways may be necessary to be activated for in vitro trans-differentiation of ADSC.
The axillary arch - morphological and histological study with clinical importance

Omar N, Abidin FZ, Das S, Abd Ghafar N, Haji Suhaimi F, Abd Latiff A, et al.

Morphologie, 2010 May;94(305):36-9.
PMID: 20359930 DOI: 10.1016/j.morpho.2010.03.001

The latissimus dorsi is a muscle of the back which forms the posterior fold of the axilla and its tendon twists to insert into the floor of the intertubercular sulcus of the humerus. Occasionally, the muscle has a muscular slip which crosses the axilla to insert into the pectoralis major. This muscular slip is often termed as "axillary arch." In the present study, we report bilateral axillary arch detected in a 45-year-old male cadaver. The average vertical length of the axillary arch measured 7 cm. The average maximum width of the uppermost, middle and lower part of the arch measured 2, 3.5 and 3.2 cm, respectively. The presence of the axillary arch is an uncommon finding in humans, considering the fact that it is solely found in the animals who prefer to hang on the trees. A histological study of the axillary arch was also performed and it showed skeletal muscle fibres which was uniformly arranged. The presence of the axillary arch may assist in the adduction of the shoulder. It may also compress the axillary vessels and nerves thereby causing resultant symptoms. Prior anatomical knowledge of the presence of axillary arch may be helpful for surgeons performing radical dissection of the axillary lymph nodes and ligation of axillary vessels, clinicians diagnosing abduction syndromes and interventional radiologists interpreting axillary mass in day to day clinical practice.
Vitamin E and factors affecting atherosclerosis in rabbits fed a cholesterol-rich diet

Ismail NM, Abdul Ghafar N, Jaarin K, Khine JH, Top GM

Int J Food Sci Nutr, 2000;51 Suppl:S79-94.
PMID: 11271860

The present study aims to examine the effects of a palm-oil-derived vitamin E mixture containing tocotrienol (approximately 70%) and tocopherol (approximately 30%) on plasma lipids and on the formation of atherosclerotic plaques in rabbits given a 2% cholesterol diet. Eighteen New Zealand White rabbits (2.2-2.8 kg) were divided into three groups; group 1 (control) was fed a normal diet, group 2 (AT) was fed a 2% cholesterol diet and group 3 (PV) was fed a 2% cholesterol diet with oral palm vitamin E (60 mg/kg body weight) given daily for 10 weeks. There were no differences in the total cholesterol and triacylglycerol levels between the AT and PV groups. The PV group had a significantly higher concentrations of HDL-c and a lower TC/HDL-c ratio compared to the AT group (P < 0.003). The aortic tissue content of cholesterol and atherosclerotic lesions were comparable in both the AT and PV groups. However, the PV group had a lower content of plasma and aortic tissue malondialdehyde (P < 0.005). Our findings suggest that despite a highly atherogenic diet, palm vitamin E improved some important plasma lipid parameters, reduced lipid peroxidation but did not have an effect on the atherosclerotic plaque formation.
Molecular concept of diabetic wound healing: effective role of herbal remedies

Soliman A, Teoh SL, Ghafar N, Das S

Mini Rev Med Chem, 2018 Oct 25.
PMID: 30360709 DOI: 10.2174/1389557518666181025155204

The incidence of diabetes mellitus (DM) is increasing worldwide. One of the main complications in DM is delayed wound healing which often requires amputation. Various drugs have been used to treat DM but they present with various complications and patients often do not comply with such treatment. This opens the door for complementary and alternative medicine. In the present review, we explore the molecular concept of wound healing occurring in different stages with special emphasis to DM. We also highlight potential herbal products such as NF3 (Chinese 2-Herb Formula), Zicao, Jing Wan Hong ointment, mixture of Adiantum capillus-veneris, Commiphora molmol, Aloe Vera, and henna, Aleo vera, Phenol-rich compound sweet gel, Jinchuang ointment, San-huang-sheng-fu (S) oil, Yi Bu A Jie extract, Astragali Radix (AR) and Rehmanniae Radix (RR), Yiqi Huayu, Tangzu yuyang ointment, Shengji Huayu recipe, Angelica sinensis, Lithospermun erythrorhison, Hippophae rhamnoides L., Curcuma longa, and Momordica charantia that could be effectively used to treat DM wounds. Future clinical trials are needed for designing potential drugs which may be effective in treating DM wounds.
Fulltext Role of MicroRNA in Proliferation Phase of Wound Healing

Soliman AM, Das S, Abd Ghafar N, Teoh SL

Front Genet, 2018;9:38.
PMID: 29491883 DOI: 10.3389/fgene.2018.00038

Wound healing is a complex biological process that is generally composed of four phases: hemostasis, inflammation, proliferation, and remodeling. The proliferation phase is crucial for effective healing compared to other phases. Many critical events occur during this phase, i.e., migration of fibroblasts, re-epithelialization, angiogenesis and wound contraction. Chronic wounds are common and are considered a major public health problem. Therefore, there is the increasing need to discover new therapeutic strategies. MicroRNA (miRNA) research in the field of wound healing is in its early phase, but the knowledge of the recent discoveries is essential for developing effective therapies for the treatment of chronic wounds. In this review, we focused on recently discovered miRNAs which are involved in the proliferation phase of wound healing in the past few years and their role in wound healing.
Fulltext Healthcare commissioners' experience with antibiotic resistance during the COVID-19 pandemic in Saudi Arabia: a qualitative study

Sulayyim HJA, Ismail R, Hamid AA, Abdul Ghafar N

J Pharm Policy Pract, 2024;17(1):2290671.
PMID: 38205192 DOI: 10.1080/20523211.2023.2290671

INTRODUCTION: The occurrence of antibiotic resistance (AR) has become a critical issue during the Novel coronavirus disease 2019 (COVID-19) pandemic. This study explores the experiences of healthcare commissioners with AR during the COVID-19 pandemic, identifies challenges, and provides recommendations for combating AR during pandemics.
METHODS: This qualitative study was multi-centered and used a phenomenological approach. Semi-structured interviews were conducted between December 2022 and January 2023 among 11 health commissioners using video calls.
RESULTS: Seven themes emerged from the data, including knowledge of AR and its consequences, the antibiotic prescription system, the future of AR and potential contributory factors, the impact of COVID-19 on AR and their relationship, the experience of AR during the COVID-19 pandemic in healthcare facilities, barriers that prevent the misuse of antibiotics during pandemics, and recommendations regarding antibiotic resistance during pandemics.
CONCLUSION: The findings of this study could be used to inform policy and practice for government healthcare workers (HCWs) and the public. Furthermore, this study identified the main challenges of AR during the pandemic, and the recommendations of health commissioners were provided accordingly. Such recommendations could be beneficial on a national and international scale to reduce the impact of future pandemics on AR.
ABBREVIATIONS: COVID-19: Novel coronavirus disease 2019; AR: Antibiotic Resistance; IPC: Infection prevention and control; MDRO: multi-drug resistant organism; ASP: Antimicrobial Stewardship Program; HCW: Healthcare worker; KSA: Kingdom of Saudi Arabia; WHO: World Health Organization; MOH: Ministry of Health; MOEWA: Ministry of Environment, Water, and Agriculture; AMR: Antimicrobial Resistance; PHCC: Primary Healthcare Center.
Fulltext Annatto-Derived Tocotrienol Promotes Mineralization of MC3T3-E1 Cells by Enhancing BMP-2 Protein Expression via Inhibiting RhoA Activation and HMG-CoA Reductase Gene Expression

Wan Hasan WN, Chin KY, Abd Ghafar N, Soelaiman IN

Drug Des Devel Ther, 2020;14:969-976.
PMID: 32184566 DOI: 10.2147/DDDT.S224941

PURPOSE: Annatto-derived tocotrienol (AnTT) has been shown to improve bone formation in animal models of osteoporosis and promote differentiation of pre-osteoblastic cells. However, the mechanism of action of AnTT in achieving these effects is unclear. This study aims to investigate the mechanism of action of AnTT on MC3T3-E1 pre-osteoblasts via the mevalonate pathway.
METHODS: Murine pre-osteoblastic cells, MC3T3-E1, were cultured with the density of 1 × 104 cells/mL and treated with 4 concentrations of AnTT (0.001-1 µg/mL). Expression of HMG-CoA reductase (HMGR) gene was carried out using qPCR after treatment with AnTT for 21 days. RhoA activation and bone morphogenetic protein-2 (BMP-2) were measured using immunoassay after 9 and 15 days of AnTT treatment. Lovastatin was used as the positive control. Mineralized nodules were detected using Von Kossa staining after 21 days of AnTT treatment.
RESULTS: The results showed that HMGR was up-regulated in the lovastatin group on day 9 and 21 compared to the control. Lovastatin also inhibited RhoA activation (day 9 and 15) and increased BMP-2 protein (day 15). On the other hand, AnTT at 0.001 μg/mL (day 3) and 0.1 μg/mL (day 21) significantly down-regulated HMGR gene expression compared to the control. On day 21, HMGR gene expression was significantly reduced in all groups compared to day 15. AnTT at 0.1 μg/mL significantly decreased RhoA activation on day 9 compared to the control. AnTT at 1 μg/mL significantly increased BMP-2 protein on day 15 compared to the control (P<0.05). Mineralized calcium nodules were more abundant in AnTT treated groups compared to the control on day 21.
CONCLUSION: AnTT suppresses the mevalonate pathway by downregulating HMGR gene expression and inhibiting RhoA activation, leading to increased BMP-2 protein in MC3T3-E1 cells. This explains the stimulating effects of AnTT on osteoblast mineralization.
Fulltext Effect of acacia honey on cultured rabbit corneal keratocytes

Ker-Woon C, Abd Ghafar N, Hui CK, Mohd Yusof YA

BMC Cell Biol., 2014;15:19.
PMID: 24885607 DOI: 10.1186/1471-2121-15-19

Acacia honey is a natural product which has proven to have therapeutic effects on skin wound healing, but its potential healing effects in corneal wound healing have not been studied. This study aimed to explore the effects of Acacia honey (AH) on corneal keratocytes morphology, proliferative capacity, cell cycle, gene and protein analyses. Keratocytes from the corneal stroma of six New Zealand white rabbits were isolated and cultured until passage 1. The optimal dose of AH in the basal medium (FD) and medium containing serum (FDS) for keratocytes proliferation was identified using MTT assay. The morphological changes, gene and protein expressions of aldehyde dehydrogenase (ALDH), marker for quiescent keratocytes and vimentin, marker for fibroblasts were detected using q-RTPCR and immunocytochemistry respectively. Flowcytometry was performed to evaluate the cell cycle analysis of corneal keratocytes.
Fulltext Effects of edible bird's nest (EBN) on cultured rabbit corneal keratocytes

Zainal Abidin F, Hui CK, Luan NS, Mohd Ramli ES, Hun LT, Abd Ghafar N

BMC Complement Altern Med, 2011;11:94.
PMID: 21992551 DOI: 10.1186/1472-6882-11-94

There has been no effective treatment or agent that is available for corneal injury in promoting corneal wound healing. Previous studies on edible bird's nest extract (EBN) had reported the presence of hormone-like substance; avian epidermal growth factor that could stimulate cell division and enhance regeneration. This study aimed to investigate the effects of EBN on corneal keratocytes proliferative capacity and phenotypical changes.
Accessory renal vessels at the upper and lower pole of the kidney: a cadaveric study with clinical implications

Hlaing KP, Das S, Sulaiman IM, Abd-Latiff A, Abd-Ghafar N, Suhaimi FH, et al.

Bratisl Lek Listy, 2010;111(5):308-10.
PMID: 20568426

The renal artery is known to exhibit variations in its number and position. The present study was performed on 50 cadaveric kidneys to observe the topographical anatomy of the accessory renal arteries (ARA) entering the upper or lower poles of the kidney. Out of 50 kidney cadaveric specimens (irrespective of sex) studied, 2 kidneys (4%) showed the presence of ARA. The presence of ARA was observed on the left and right kidneys, respectively. In one left kidney, we observed in addition to the usual renal artery, an ARA near the lower pole of the kidney which divided into anterior and posterior branches. Another right kidney specimen exhibited the presence of single and double ARA at the upper and the lower poles, respectively. The presence of ARA, both at the upper and lower poles is a rare entity. No medical history of the cadavers was available to corroborate the clinical findings. Additional renal vessels may signify a developmental defect. Anatomical knowledge of the variations in the renal vascular supply may be important for abdominal imaging studies and surgical operations involving renal transplantations. The present study discusses in detail the anatomical features and clinical implications of ARA located at both the upper and lower poles of the kidney (Fig. 2, Ref. 15). Full Text (Free, PDF) www.bmj.sk.
Atlanto-occipital fusion: an osteological study with clinical implications

Kassim NM, Latiff AA, Das S, Ghafar NA, Suhaimi FH, Othman F, et al.

Bratisl Lek Listy, 2010;111(10):562-5.
PMID: 21125803

Atlanto-occipital fusion may be symptomatic or asymptomatic in nature. The anomaly may be incidentally detected at autopsies or during routine cadaveric dissections. The fusion of the atlas with occipital bone may result in the compression of vertebral artery and first cervical nerve.
Absence of the peroneus tertius muscle: cadaveric study with clinical considerations

Das S, Haji Suhaimi F, Abd Latiff A, Pa Pa Hlaing K, Abd Ghafar N, Othman F

Rom J Morphol Embryol, 2009;50(3):509-11.
PMID: 19690784

Peroneus tertius (PT) is a muscle of the anterior compartment of the leg. The PT muscle originates from the anterior surface of the fibula and the interosseous membrane and inserts into the medial side of the dorsal region of the fifth metatarsal bone. During routine dissection, we observed the absence of PT on the left lower limb of a cadaver. Usually, the PT is involved in dorsiflexion and eversion of the foot. In many cases, the absence of PT maybe asymptomatic and it may be incidentally detected during cadaveric dissections or autopsies. The existence of PT may help in the swing phase of bipedal walking. The PT may be used for tendon graft surgeries. The pull of the PT may be responsible for causing stress on the fifth metacarpal and account for all stress fractures in any individual. The absence of the PT may puzzle any transplant and foot surgeons performing graft operations. We as anatomists discuss the clinical implications of the absence of PT.
Ex vivo growth of rabbit bulbar, fornix and palpebral conjunctival epithelia in a serum-free and feeder layer-free culture system

Nizam MH, Ruszymah BH, Chua KH, Ghafar NA, Hamzah JC

Med J Malaysia, 2008 Jul;63 Suppl A:111-2.
PMID: 19025010

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.
Scanning electron microscopy of cornea re-epithelization after transplanted with bilayered corneal construct

Masrudin SS, Ghafar NA, Saidi M, Aminuddin BS, Rahmat A, Ruszymah BH, et al.

Med J Malaysia, 2008 Jul;63 Suppl A:109-10.
PMID: 19025009

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.
Fulltext Gelam honey potentiates ex vivo corneal keratocytes proliferation with desirable phenotype expression

Yusof AM, Abd Ghafar N, Kamarudin TA, Hui CK, Yusof YA

BMC Complement Altern Med, 2016;16:76.
PMID: 26912349 DOI: 10.1186/s12906-016-1055-7

This study aimed to evaluate the effects of Gelam honey on corneal keratocytes proliferative capacity and phenotypic characterization via MTT assay, gene expression and immunocytochemistry.
Fulltext The effects of acacia honey on in vitro corneal abrasion wound healing model

Ker-Woon C, Abd Ghafar N, Hui CK, Mohd Yusof YA, Wan Ngah WZ

BMC Cell Biol., 2015;16:2.
PMID: 25887200 DOI: 10.1186/s12860-015-0053-9

Acacia honey (AH) has been proven to improve skin wound healing, but its therapeutic effects on corneal epithelium has not been elucidated to date. This study aimed to investigate the effects of AH on cultured corneal epithelial cells (CEC) on in vitro corneal abrasion wound healing model. Six New Zealand white rabbits' CEC were isolated and cultured until passage 1. Circular wound area was created onto a confluent monolayer CEC using a corneal trephine which mimicked corneal abrasion and treated with 0.025% AH supplemented in basal medium (BM) and complete cornea medium (CCM). Wound healing was measured as the percentage of wound closure by the migration of CEC on day 0, day 3 and day 6, post wound creation. The morphological changes of CEC were assessed via phase contrast microscopy. Gene and protein expressions of cytokeratin (CK3), fibronectin and cluster of differentiation 44 (CD44) in AH treated groups and control groups were determined by real-time PCR and immunocytochemistry, respectively.
The role of Gelam honey in accelerating reepithelialization of ex vivo corneal abrasion model

Azmi MF, Abd Ghafar N, Che Hamzah J, Chua KH, Ng SL

J Food Biochem, 2021 04;45(4):e13645.
PMID: 33569805 DOI: 10.1111/jfbc.13645

This study aimed to investigate the role of Gelam honey (GH) in accelerating reepithelialization of corneal abrasion. Corneal epithelial cells (CEC) isolated from New Zealand white rabbit corneas, were cultured and circular-shaped wounds were created onto them, representing the corneal abrasion model. These wounds were treated with basal (BM) and cornea media (CM) supplemented with GH. The percentage of wound closure was measured on day 0, 3, and 5. Expressions of cytokeratin 3 (CK3), cluster of differentiation 44 (CD 44), and connexin 43 (Cx43) were analyzed via qRT-PCR and immunocytochemistry. The results showed CEC cultured in GH-enriched media reepithelialized faster compared to control. Corneal abrasion treated with CM supplemented with GH closed completely on day 5. CK3, CD44, and Cx43 expressions correspond to the stages of reepithelialization. In conclusion, GH promotes the healing of the ex vivo corneal abrasion model. Further explorations of its potential as adjuvant therapy in treating corneal injuries are needed. PRACTICAL APPLICATIONS: Honey has been reported to have many medicinal properties including antibacterial, anti-inflammatory, and the ability to promote skin wound healing. However, the effects of honey on corneal wound healing have not been fully elucidated. In the present study, we aimed to determine the effects of Gelam honey (GH), well-known local honey obtained from the beehive of Gelam trees (Melaleuca spp.), on the ex vivo corneal abrasion model via cell migration study and analysis of genes and proteins during corneal epithelial wound healing. GH has proven to have accelerated effects on the corneal epithelial cell migration during the closure of the ex vivo corneal abrasion wound model. The expressions of the genes and proteins of the corneal epithelial wound healing markers were in accordance with the stages of healing. Therefore, GH has the potential to be developed as adjuvant therapy in the form of GH-based eye drop in treating corneal injuries.
Fulltext Acacia honey accelerates in vitro corneal ulcer wound healing model

Abd Ghafar N, Ker-Woon C, Hui CK, Mohd Yusof YA, Wan Ngah WZ

BMC Complement Altern Med, 2016 Jul 29;16:259.
PMID: 27473120 DOI: 10.1186/s12906-016-1248-0

BACKGROUND: The study aimed to evaluate the effects of Acacia honey (AH) on the migration, differentiation and healing properties of the cultured rabbit corneal fibroblasts.
METHODS: Stromal derived corneal fibroblasts from New Zealand White rabbit (n = 6) were isolated and cultured until passage 1. In vitro corneal ulcer was created using a 4 mm corneal trephine onto confluent cultures and treated with basal medium (FD), medium containing serum (FDS), with and without 0.025 % AH. Wound areas were recorded at day 0, 3 and 6 post wound creation. Genes and proteins associated with wound healing and differentiation such as aldehyde dehydrogenase (ALDH), vimentin, alpha-smooth muscle actin (α-SMA), collagen type I, lumican and matrix metalloproteinase 12 (MMP12) were evaluated using qRT-PCR and immunocytochemistry respectively.
RESULTS: Cells cultured with AH-enriched FDS media achieved complete wound closure at day 6 post wound creation. The cells cultured in AH-enriched FDS media increased the expression of vimentin, collagen type I and lumican genes and decreased the ALDH, α-SMA and MMP12 gene expressions. Protein expression of ALDH, vimentin and α-SMA were in accordance with the gene expression analyses.
CONCLUSION: These results demonstrated AH accelerate corneal fibroblasts migration and differentiation of the in vitro corneal ulcer model while increasing the genes and proteins associated with stromal wound healing.

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