A new invention, papain-based wound cleanser is formulated by incorporating papain, a proteolytic enzyme extracted from Carica papaya into the formulation. This cleanser is invented to simplify the methods in wound management by combining wound cleansing and wound debridement using a single formulation. This study describes the preparation and preclinical study of papain-based wound cleanser in accelerating wound healing. In this study, papain-based wound cleanser was used to treat wound incision on Sprague-Dawley rats while distilled water and Betadine were used as negative and positive control. Twenty-seven clinically healthy white rats were randomly divided into three groups and treated accordingly until the 21st day post-incision. Wound reduction rates and histological analysis were obtained to asses the healing pattern. Rats treated with papain-based wound cleanser showed a progressive wound healing based on the wound reduction rates and histological analysis when compared with rats treated with distilled water and Betadine. Better collagen deposition and presence of skin organelles in rats treated with papain-based wound cleanser demonstrated its efficacy in promoting wound healing. In addition to its wound healing effect, papain-based wound cleanser is also integrated with antibacterial properties which make it a complete package for wound management. However, further studies should be carried out to ensure its safety for human usage.
An efficient in vitro plant regeneration system was established for elite, recalcitrant Malaysian indica rice, Oryza sativa L. CV. MR 219 using mature seeds as explant on Murashige and Skoog and Chu N6 media containing 2,4-dichlorophenoxy acetic acid and kinetin either alone or in different combinations. L-proline, casein hydrolysate and L-glutamine were added to callus induction media for enhancement of embryogenic callus induction. The highest frequency of friable callus induction (84%) was observed in N6 medium containing 2.5 mg l(-1) 2,4-dichlorophenoxy acetic acid, 0.2 mg l(-1) kinetin, 2.5 mg l(-1) L-proline, 300 mg l(-1) casein hydrolysate, 20 mg l(-1) L-glutamine and 30 g l(-1) sucrose under culture in continuous lighting conditions. The maximum regeneration frequency (71%) was observed, when 30-day-old N6 friable calli were cultured on MS medium supplemented with 3 mg l(-1) 6-benzyl aminopurine, 1 mg l(-1) naphthalene acetic acid, 2.5 mg l(-1) L-proline, 300 mg l(-1) casein hydrolysate and 3% maltose. Developed shoots were rooted in half strength MS medium supplemented with 2% sucrose and were successfully transplanted to soil with 95% survival. This protocol may be used for other recalcitrant indica rice genotypes and to transfer desirable genes in to Malaysian indica rice cultivar MR219 for crop improvement.
The objective of this study was to regenerate the tracheal epithelium using autologous nasal respiratory epithelial cells in a sheep model. Respiratory epithelium and fibroblast cells were harvested from nasal turbinates and cultured for 1 week. After confluence, respiratory epithelium and fibroblast cells were suspended in autologous fibrin polymerized separately to form a tissue-engineered respiratory epithelial construct (TEREC). A 3 × 2 cm² tracheal mucosal defect was created, and implanted with TEREC and titanium mesh as a temporary scaffold. The control groups were divided into 2 groups: polymerized autologous fibrin devoid of cells (group 1), and no construct implanted (group 2). All sheep were euthanized at 4 weeks of implantation. Gross observation of the trachea showed minimal luminal stenosis formation in the experimental group compared to the control groups. Macroscopic evaluation revealed significant mucosal fibrosis in control group 1 (71.8%) as compared to the experimental group (7%). Hematoxylin and eosin staining revealed the presence of minimal overgrowth of fibrous connective tissue covered by respiratory epithelium. A positive red fluorescence staining of PKH26 on engineered tissue 4 weeks after implantation confirmed the presence of cultured nasal respiratory epithelial cells intercalated with native tracheal epithelial cells. Scanning electron microscopy showed the presence of short microvilli representing immature cilia on the surface of the epithelium. Our study showed that TEREC was a good replacement for a tracheal mucosal defect and was able to promote natural regenesis of the tracheal epithelium with minimal fibrosis. This study highlighted a new technique in the treatment of tracheal stenosis.
Tissue regeneration is a rapidly growing field providing a beacon of hope in the field of restorative and endodontics. Root canal treatment involves the removal of pulp tissue and replacement by an inorganic materials where as regenerative endodontics deals with replacement with healthy pulp to revitalize the teeth .Research in the field of tissue engineering and material science have lead to significant progress but still is plague with lots of drawbacks and failures, hence it is still not being adapted as routine clinical procedures .The purpose of this article is to review the advances made in regenerative endodontics and the future scopes.
PURPOSE: The purpose of this study was to evaluate the quality of articular cartilage regeneration after arthroscopic subchondral drilling followed by postoperative intraarticular injections of autologous peripheral blood progenitor cells (PBPCs) in combination with hyaluronic acid (HA).
METHODS: Five patients underwent second-look arthroscopy with chondral core biopsy. These 5 patients are part of a larger pilot study in which 180 patients with International Cartilage Repair Society grade III and IV lesions of the knee joint underwent arthroscopic subchondral drilling followed by postoperative intra-articular injections. Continuous passive motion was used on the operated knee 2 hours per day for 4 weeks. Partial weight bearing was observed for the first 6 to 8 weeks. Autologous PBPCs were harvested 1 week after surgery. One week after surgery, 8 mL of the harvested PBPCs in combination with 2 mL of HA was injected intra-articularly into the operated knee. The remaining PBPCs were divided into vials and cryopreserved. A total of 5 weekly intra-articular injections were given.
RESULTS: Second-look arthroscopy confirmed articular cartilage regeneration, and histologic sections showed features of hyaline cartilage. Apart from the minimal discomfort of PBPC harvesting and localized pain associated with the intra-articular injections, there were no other notable adverse reactions.
CONCLUSIONS: Articular hyaline cartilage regeneration is possible with arthroscopic subchondral drilling followed by postoperative intraarticular injections of autologous PBPCs in combination with HA.
LEVEL OF EVIDENCE: Level IV, therapeutic case series.
The staggering number of publications featuring the use of stem cells has revolutionized regenerative medicine research. Preclinical studies indicate that allogeneic human mesenchymal stem cells (MSCs) may be useful for the treatment of several clinical disorders, including sepsis, acute renal failure, acute myocardial infarction, and more recently, acute lung injury (ALI). However, considerable success would not be obtained in clinical trials due to poor survival of transplanted cells under the influence of inflammatory conditions. Despite robust approaches like cellular reprogramming, scaffolds and conditioned media have been tested to overcome this problem; however the success rate of these approaches remain questionable. Recently, pretreatment of bioactive compounds in vitro have been shown to suppress cell apoptosis and promote cell survival. Quite likely a similar phenomenon can take place in vivo. Based on such studies, we hypothesize that MSCs derived from human post-natal tissues could be conditioned and prepared for targeted disease therapy. Depending on the disease condition, the MSCs could be treated prior to delivery with appropriate bioactive compounds to allow them survive longer and perform a better role as biocatalyst. The advantage of this approach could be the tailor made availability of MSCs preconditioned with appropriate bioactive compounds for disease specific therapy. Therefore, the choice of suitable bioactive molecule is likely to enhance the efficacy of targeted stem cell therapy and preconditioning may provide a novel strategy in maximizing biological and functional properties of MSCs.
Murraya koenigii (Linn.) Spreng (curry leaf) is widely used as a nephroprotective agent in kidney's infirmities among diabetics by the traditional practitioners in Malaysia. However, the latter role of curry leaf has been grossly under reported and is yet to receive proper scientific evaluation.
Cell-based therapy using autologous cells has been suggested as a potential approach for periodontal tissue regeneration. Spheroid systems are a form of three-dimensional cell culture that promotes cell matrix interaction, which could recapitulate the aspect of cell homeostasis in vivo. The aim of this study is to assess the interaction of periodontal fibroblast spheroids with synthetic and collagen-based membranes that have been used in guided tissue regeneration.
Nowadays, many researches were conducted in minimizing tissue culture technology due to the overhead of cost needed. The purpose of this study was to investigate the effects of using five kinds of organic additives at four level concentrations responsive to the number of shoots produced for eight weeks in culture. Stem segment explants of Celosia sp. were cultured on MS medium that have been supplemented with different kinds of extract juice that serve as organic additives which are mature coconut, young coconut, papaya, banana and tomato at 20, 30, 50 and 70 ml L-1. The numbers of shoot on each explant were recorded and the mean of ten replicates explants were calculated. Among the media used, young coconut water at 70 ml L1- induced the highest shoot regeneration (14.21+/-8.26), followed by mature coconut water at 50 ml L-1 (13.14+/-10.33). Banana and tomato juice promote highest shoot regeneration of stem segments at 50 ml L-1 that produced 9.57+/-4.68 and 9.28+/-5.82 shoots per explants, respectively. While the lowest concentration which at 20 ml L-1 of papaya juice showed highest shoot regeneration (10.50+/-3.45) produced among the three other concentration tested. Statistical results showed that there were significant differences interactions effects (p<0.05) in terms of number of shoot regenerated between the types of extracts juices determined by ANOVA test. Comparing number of shoots regenerated that were cultured in control media, it showed higher than all of experimental medium composition. There were no big different in cost required in preparation of control media and the experimental media. Applications of five kinds of local fruit in tissue culture media should be considered since it responsive in shoot regeneration.
We have investigated the effect of three potential scar-reducing agents applied at a sciatic nerve repair site in C57-black-6 mice. Under anaesthesia the nerve was transected, repaired using four epineurial sutures, and 100 μl of either triamcinolone acetonide (1 mg/100 μl), an interleukin-10 peptide fragment (125 ng/100 μl or 500 ng/100 μl) or mannose-6-phosphate (M6P, 200 mM or 600 mM) was injected into and around the nerve. After 6 weeks the extent of regeneration was assessed electrophysiologically by determining the ratio of the compound action potential (CAP) modulus evoked by electrical stimulation of the nerve 2 mm distal or proximal to the repair site. The conduction velocity of the fastest components in the CAP was also calculated. The percentage area of collagen staining (PAS) at the repair site was analysed using Picrosirius Red and image analysis. Comparisons were made with a placebo group (100 μl of phosphate buffered saline) and sham-operated controls. The median CAP modulus ratio in the 600 mM M6P group was 0.44, which was significantly higher than in the placebo group (0.24, P=0.012: Kruskal-Wallis test). Conduction velocities were also faster in the 600 mM M6P group (median 30 m s(-1)) than in the placebo group (median 27.8 m s(-1); P=0.0197: Kruskal-Wallis test). None of the other treated groups were significantly different from the placebo, and all had significantly lower CAP ratios than the sham controls (P<0.05). All repair groups had a significantly higher PAS for collagen than sham controls. We conclude that the administration of 600 mM mannose-6-phosphate to a nerve repair site enhances axonal regeneration.
Spinal cord injuries (SCIs) are a common form of trauma that leaves a huge trail of morbidity and human suffering in its wake. They occur mostly among the young, causing severe physical, psychological, social and economic burdens. The treatment of this condition has rather been disappointing; most of the management strategies being mainly supportive and prophylactic. In recent years there has been an emerging interest in the use of stem cells to regenerate the nervous tissue that has been damaged or lost. Although there has been much hype and unfounded hope, modest successes have been witnessed, and it is possible that these therapeutic strategies may have much more to offer in the future. This paper will review the current strategies of exploring cell-based therapies, mainly different types of stem cells to treat SCI along with the evidence that has been accumulated over the past decade in a rational bench-to-bedside approach. Furthermore, critical aspects such as the mode of delivery and ethical considerations are also discussed along with feasible suggestions for future translational research to provide a contextual picture of the current state of advancements in this field. The impediments to regeneration in the site of injury are briefly explained along with the benefits and drawbacks of different cell types used in the treatment of this condition. We hope that this review will offer a significant insight into this challenging clinical condition.
The use of bone grafts in treating non- or delayed unions as the result of large bone loss is well established. However, despite good outcomes, the time to achieve complete union is still considerably long. To overcome this problem, the use of platelet-rich plasma (PRP) has been advocated albeit with varying success. To determine the true effectiveness of PRP in treating non-/delayed unions, a study was conducted using (n=12) rabbit models.
Understanding the fundamental processes involved in repairing the airway wall following injury is fundamental to understanding the way in which these processes are perturbed during disease pathology. Indeed complex diseases such as asthma and chronic obstructive pulmonary disease (COPD) have at their core evidence of airway wall remodeling processes that play a crucial functional role in these diseases. The authors sought to understand the dynamic cellular events that occur during bronchial airway epithelial repair in sheep. The injury was induced by endobronchial brush biopsy (BBr), a process that causes epithelial débridement and induces a consequential repair process. In addition, the current experimental protocol allowed for the time-dependent changes in airway wall morphology to be studied both within and between animals. The initial débridement was followed by evidence of dedifferentiation in the intact epithelium at the wound margins, followed by proliferation of cells both within the epithelium and in the deeper wall structures, notably in association with the submucosal glands and smooth muscle bundles. Seven days after injury, although the airway wall was thickened at the site of damage, the epithelial layer was intact, with evidence of redifferentiation. These studies, in demonstrating broad agreement with previous studies in small animals, indicate the wider relevance of this system as a comparative model and should provide a solid basis upon which to further characterize the critical cellular and molecular interactions that underlie both effective restitution and pathological repair.
Nerve crush injury is a well-established axonotmetic model in experimental regeneration studies to investigate the impact of various pharmacological treatments. Hericium erinaceus is a temperate mushroom but is now being cultivated in tropical Malaysia. In this study, we investigated the activity of aqueous extract of H. erinaceus fresh fruiting bodies in promoting functional recovery following an axonotmetic peroneal nerve injury in adult female Sprague-Dawley rats by daily oral administration. The aim was to investigate the possible use of this mushroom in the treatment of injured nerve. Functional recovery was assessed in behavioral experiment by walking track analysis. Peroneal functional index (PFI) was determined before surgery and after surgery as rats showed signs of recovery. Histological examinations were performed on peroneal nerve by immunofluorescence staining and neuromuscular junction by combined silver-cholinesterase stain. Analysis of PFI indicated that return of hind limb function occurred earlier in rats of aqueous extract or mecobalamin (positive control) group compared to negative control group. Regeneration of axons and reinnervation of motor endplates in extensor digitorum longus muscle in rats of aqueous extract or mecobalamin group developed better than in negative control group. These data suggest that daily oral administration of aqueous extract of H. erinaceus fresh fruiting bodies could promote the regeneration of injured rat peroneal nerve in the early stage of recovery.
The haruan (Channa striatus) is an indigenous, predatory freshwater fish of Malaysia. It is a common food fish among the local populace with traditionally identified pharmacological benefits in treating wound and pain and in boosting energy of the sick. Channa striatus is also a subject of renewed interest in Malaysian folk medicine in the search for a better cure for diseases and ailments. Amino acids and fatty acids, found in high concentrations in the fish, might have contributed to its pharmacological properties. Important amino acids of the fish include glycine, lysine and arginine, while its fatty acids are arachidonic acid, palmitic acid and docosahexaenoic acid. They appear to effect their influence through the formation of several types of bioactive molecules. Extracts of the fish are produced from whole fish, roe, mucus and skin of the fish. This review updates research findings on potential uses of Channa striatus, beyond the traditional prescription as a wound healer, pain reliever and energy booster to include its properties as a ACE-inhibitor, anti-depressant and neuroregenerative agent. The fish appears to have wide-ranging medical uses and should be studied more intensively to unearth its other properties and mechanisms of action.
This review summarises the major developments of macroporous bioceramics used mainly for repairing bone defects. Porous bioceramics have been receiving attention ever since their larger surface area was reported to be beneficial for the formation of more rigid bonds with host tissues. The study of porous bioceramics is important to overcome the less favourable bonds formed between dense bioceramics and host tissues, especially in healing bone defects. Macroporous bioceramics, which have been studied extensively, include hydroxyapatite, tricalcium phosphate, alumina, and zirconia. The pore size and interconnections both have significant effects on the growth rate of bone tissues. The optimum pore size of hydroxyapatite scaffolds for bone growth was found to be 300 µm. The existence of interconnections between pores is critical during the initial stage of tissue ingrowth on porous hydroxyapatite scaffolds. Furthermore, pore formation on β-tricalcium phosphate scaffolds also allowed the impregnation of growth factors and cells to improve bone tissues growth significantly. The formation of vascularised tissues was observed on macroporous alumina but did not take place in the case of dense alumina due to its bioinert nature. A macroporous alumina coating on scaffolds was able to improve the overall mechanical properties, and it enabled the impregnation of bioactive materials that could increase the bone growth rate. Despite the bioinertness of zirconia, porous zirconia was useful in designing scaffolds with superior mechanical properties after being coated with bioactive materials. The pores in zirconia were believed to improve the bone growth on the coated system. In summary, although the formation of pores in bioceramics may adversely affect mechanical properties, the advantages provided by the pores are crucial in repairing bone defects.
Orthodontic elastic bands are an important iatrogenic etiologic factor in the causation of periodontal attachment apparatus breakdown. Appropriate diagnosis and a well constructed treatment plan tailor-made to suit the requirements of the particular patient is imperative for management of periodontal lesions induced by subgingival retention of rubber band. There are conflicting reports regarding the reattachment and regeneration of lost periodontal supporting tissues in such cases. The present case report highlights the spontaneous reversal and correction of periodontal destruction due to iatrogenic orthodontic elastic band displacement deep into the subgingival tissues.
Tissue culture studies of Celosia cristata were established from various explants and the effects of various hormones on morphogenesis of this species were examined. It was found that complete plant regeneration occurred at highest percentage on MS medium supplemented with 2.0 mg/L NAA and 1.5 mg/L BAP, with the best response showed by shoot explants. In vitro flowering was observed on MS basal medium after six weeks. The occurrence of somaclonal variation and changes in cellular behavior from in vivo and in vitro grown plants were investigated through cytological studies and image analysis. It was observed that Mitotic Index (MI), mean chromosome numbers, and mean nuclear to cell area ratio of in vitro root meristem cells were slightly higher compared to in vivo values. However, in vitro plants produced lower mean cell areas but higher nuclear areas when compared to in vivo plants. Thus, no occurrence of somaclonal variation was detected, and this was supported by morphological features of the in vitro plants.
Understanding the mechanisms underlying the process of regeneration and repair of airway epithelial structures demands close characterization of the associated cellular and molecular events. The choice of an animal model system to study these processes and the role of lung stem cells is debatable since ideally the chosen animal model should offer a valid comparison with the human lung. Species differences may include the complex three-dimensional lung structures, cellular composition of the lung airway as well as transcriptional control of the molecular events in response to airway epithelium regeneration, and repair following injury. In this paper, we discuss issues related to the study of the lung repair and regeneration including the role of putative stem cells in small- and large-animal models. At the end of this paper, the author discuss the potential for using sheep as a model which can help bridge the gap between small-animal model systems and humans.
Different concentrations of N6-benzylaminopurine (BAP) and indole acetic acid (IAA) in Murashige and Skoog based medium were assessed for their effects on shoot multiplication, nodule-like meristem proliferation and plant regeneration of the Malaysian banana cultivars Pisang Mas, Pisang Nangka, Pisang Berangan and Pisang Awak. BAP at 1-14 mg L-1 with or without 0.2 mg L-1 IAA, or BAP at 7-14 mg L-1with the same concentration of IAA, was evaluated for shoot multiplication from shoot tips and the proliferation of nodule-like meristems from scalps, respectively. Plant regeneration from scalps was assessed using 1 mg L-1BAP and 0.2 mg L-1 IAA separately, or a combination of these two growth regulators. Data on shoot multiplication, the proliferation of nodule-like meristems with associated plant regeneration were recorded after 30 days of culture. A maximum of 5 shoots per original shoot tip was achieved on medium supplemented with BAP at 5 mg L-1 (Pisang Nangka), 6 mg L-1(Pisang Mas and Pisang Berangan), or 7 mg L-1 (Pisang Awak), with 0.2 mg L-1 IAA. BAP at 11 mg L-1 with 0.2 mg L-1 IAA induced the most highly proliferating nodule-like meristems in the four banana cultivars. Plant regeneration from scalps was optimum in all cases on medium containing 1 mg L-1 BAP and 0.2 mg L-1 IAA. This is the first report on the successful induction of highly proliferating nodule-like meristems and plant regeneration from scalps of the Malaysian banana cultivars Pisang Mas, Pisang Nangka, Pisang Berangan and Pisang Awak.