Efficacy of autologous bone marrow derived Mesenchymal stem cells (MSCs), osteoblasts and osteoblasts derived exosome in the reversal of ovariectomy (OVX) induced osteoporosis in rabbit model.

Background: A recent study showed that OVX-induced osteoporosis was reversed after injection of osteoblasts cultured from the bone marrow in rats. The present study evaluated the effect of injecting MSCs, osteoblasts, and exosomes isolated from osteoblasts for the treatment of osteoporosis in the rabbit model. Methods: Osteoporosis was created in 40 rabbits by performing ovareictomy at 6 months of age, and 1 mg/kg body weight of methyl prednisolone sodium succinate was injected daily for 8 weeks. Animals were fed twice daily and were given water ad libitum. MSCs and osteoblasts were grown from the bone marrow as per the methodology described earlier. From osteoblasts, exosomes were extracted. After the 15th day, MSCs (Group 2), osteoblasts (Group 3), and exosomes (Group 4) were injected into 5 animals each, and 0.5 ml of normal saline were injected into the control group (Group 1). After 12 weeks (11 months of age), all the animals were euthanized. The whole femur and the lumbar vertebrae 3-5 were dissected out and were subjected to radiological assessment using high-resolution peripheral quantitative computerized tomography (HRpQCT). All parameters of the bone volume, trabecular number, thickness, and spacing were assessed using SPSS (Statistical Package for the Social Sciences), version 21.0, Chicago, Illinois. A p value of <0.05 was considered Statistically significant with a confidence interval (CI) of 95%. Results: Structural indices of the osteoblasts-injected animals were significantly better than the control group for the distal femur. The most significant improvement was seen in the osteoblasts, MSCs, and exosomes group in that order. The p value of all parameters was <0.0001 in the osteoblasts group, whereas the total and bone volume had a lower p value in the MSCs group. In the osteoblasts group, the positive changes were similar in the distal femur and lumbar vertebrae, but with MSCs and exosomes, the changes were more pronounced in the vertebral spine than the distal femur. Conclusions: This study shows that autologous bone marrow-derived osteoblasts have the robust influence of reversing OVX-induced osteoporosis in rabbits.

Spinal cord injury regeneration using autologous bone marrow-derived neurocytes and rat embryonic stem cells: A comparative study in rats.

BACKGROUND: Spinal cord injury (SCI) is an important cause of traumatic paralysis and is mainly due to motor vehicle accidents. However, there is no definite treatment for spinal cord damage. AIM: To assess the outcome of rat embryonic stem cells (rESC) and autologous bone marrow-derived neurocytes (ABMDN) treatment in iatrogenic SCI created in rats, and to compare the efficacy of the two different cell types. METHODS: The study comprised 45 male Wistar rats weighing between 250 and 300 g, which were divided into three groups, the control, rESC and ABMDN groups. The anesthetized animals underwent exposure of the thoracic 8th to lumbar 1st vertebrae. A T10-thoracic 12th vertebrae laminectomy was performed to expose the spinal cord. A drop-weight injury using a 10 g weight from a height of 25 cm onto the exposed spinal cord was conducted. The wound was closed in layers. The urinary bladder was manually evacuated twice daily and after each evacuation Ringer lactate 5 mL/100 g was administered, twice daily after each bladder evacuation for the first 7 postoperative days. On the 10th day, the rats underwent nerve conduction studies and behavioral assessment [Basso, Beattie, Brenham (BBB)] to confirm paraplegia. Rat embryonic stem cells, ABMDN and saline were injected on the 10th day. The animals were euthanized after 8 wk and the spinal cord was isolated, removed and placed in 2% formalin for histopathological analysis to assess the healing of neural tissues at the axonal level. RESULTS: All the animals tolerated the procedure well. The BBB scale scoring showed that at the end of the first week no recovery was observed in the groups. Post-injection, there was a strong and significant improvement in rats receiving rESC and ABMDN as compared to the control group based on the BBB scale, and the Train-of-four-Watch SX acceleromyography device exhibited statistically significant (P < 0.0001) regeneration of neural tissue after SCI. Histological evaluation of the spinal cord showed maximum vacuolization and least gliosis in the control group compared to the rESC and ABMDN treated animals. In the ABMDN group, limited vacuolization and more prominent gliosis were observed in all specimens as compared to the control and rESC groups. CONCLUSION: This study provided strong evidence to support that transplantation of rESC and ABMDN can improve functional recovery after iatrogenic SCI. The transplanted cells showed a beneficial therapeutic effect when compared to the control group.

Stem-cell therapy for ovariectomy-induced osteoporosis in rats: a comparison of three treatment modalities.

Background: Recent studies have shown that ovariectomy-induced osteoporosis in rats can be reversed by infusion of osteoblasts cultured from mesenchymal stem cells (MSCs). This study compares the influence of MSCs, osteoblasts, and exosomes derived from osteoblasts for the treatment of osteoporosis. Methods: Osteoporosis was induced in 40 female Sprague Dawley rats by performing ovariectomy. After 12 weeks, bone marrow was harvested and MSCs separated from bone-marrow aspirate as described by Piao et al. After 15 days, autologous osteogenically differentiated cells from the MSCs were available. Exosomes were isolated from osteoblasts by modification of the technique described by Ge et al. MSCs and osteoblasts (106 cells in 0.5 mL normal saline) and exosomes (100 µg protein) were injected into the tail veins of the animals. Animals were euthanized after 12 weeks and femurs and lumbar spines dissected and analyzed using high-resolution peripheral quantitative computed tomography. Results: When compared to the control group, osteoblast-treated animals showed significant differences in all parameters compared, with P-values ranging between <0.002 and <0.0001. Comparison among osteoblasts, MSCs, and exosomes, showed that osteoblasts had positive and statistically significant new-bone formation. The comparison for the spine was similar to the distal femur for osteoblasts. Conclusion: This study showed robust positive bone-forming changes after osteoblast injection in the distal femur and the spine when compared to controls, MSCs, and exosomes.

Topical vitamin D3: A randomized controlled trial (RCT).

OBJECTIVE: The intent of this study was to test the effect of Top-D, a topical Vitamin D preparation, in delivering vitamin D. METHODS: Five hundred and fifty healthy patients, with vitamin D insufficiency and deficiency were recruited after written informed consent. Demographic data was recorded, adequate history and clinical examination was done to rule out any metabolic diseases. Complete blood picture, serum calcium, phosphorous, Parathormone and 25 Hydroxy-vitamin D3 (25OHD) was carried out before enrollment of the patients. Patients were divided randomly into two groups 350 in study group and 200 in the control group. Patients in the study group were given Top-D (Vitamin D3 gel made from proniosomal technology) to apply daily on the skin. Top-D 1 g contained 5000 IU of vitamin D3. The control group was given 1 g of Aloe vera gel to be applied every day. The two groups had no knowledge to which group they belong. After 4 months serum 25OHD was tested again. RESULTS: Three hundred and forty five patients in study group and 192 in control group completed the study. The mean age of the patients in the both the groups was 42 years (18-80 years). The pretreatment 25OHD level in the study group was 11.03 ± 4.57 (2-12) ng/l compared to the control group 10.36 ± 4.09 (2-21) and post treatment the levels were 37.17 ± 6.04 (12-54) ng/ml and 10.51 ± 3.5 (2-19) ng/ml (p < 0.001). CONCLUSION: The results of this study indicate that transdermal route of vitamin D is potentially, safe and can give desired results to raise the vitamin D levels. This route is an alternate route for supplementation of vitamin D which should be utilized.

Bone Marrow-derived Osteoblasts in the Management of Ovariectomy induced Osteoporosis in Rats.

Background: The objective of this study is to assess if infusion of osteoblasts can temporarily reverse osteoporosis in rats. Methods: Osteoporosis was induced in 20 female Sprague-Dawley rats by performing ovariectomy (OVX) that was carried out at 4 weeks of age. At 3 months a biopsy of the iliac crest was made to assess the bone quality and the same site bone marrow was harvested. From the bone marrow aspirate, MSCs were separated. Osteoblasts were then generated and were characterized using Alizarin red staining. Osteoblasts were injected in the tail vein of 10 rats. Two weeks after the injection of osteoblasts, a second biopsy was done. Animals were euthanized after 8 weeks of osteoblasts infusion by overdose of ketamine mixed with xylazine. The whole femurs and lumbar spine were dissected and the specimens were stored in 2% formalin. The specimens were analyzed using HRpQCT (High-resolution peripheral quantitative computerized tomography (µCT 100, SCANCO Medical AG, Brüttisellen, Switzerland). Results: In all the 10 animals from which bone aspiration was performed, osteoblasts were cultured and transplanted. Analysis showed that there was significant bone formation at bone sites of distal femur and lumbar spine (<0.001), with increased number of trabeculae and thickness (P<0.001). Further analysis revealed that there was robust bone formation in the animals that had osteoblasts injection. Conclusions: This preliminary study indicates that osteoblasts infusion can lead to new bone formation in osteoporosis induced by ovariectomy in rats.

Morbidity and mortality after fragility hip fracture in a Saudi Arabian population: Report from a single center.

Objective To determine the functional morbidity and mortality after fragility hip fracture and compare the mortality with three other common diseases. Methods Data were collected from patients admitted to King Fahd Hospital of the University, AlKhobar from January 2010 to December 2014. Demographic data included the preoperative American Society of Anesthesiologists (ASA) score as assessed by the anesthetist and the type of surgery. Personal and telephone interviews were performed, and data were entered into a database and analyzed. Results We identified 203 patients with fragility proximal femoral fractures, and the data of 189 patients (109 male, 80 female; average age, 66.90 ± 13.43 years) were available for analysis. The overall mortality rate was 26.98% (51 patients). The mortality rate was significantly higher among patients with an ASA score of 4 (36.36%) than 1 (20.45%). With respect to morbidity, only 48.23% of patients were able to return to their pre-fracture status; 32.35% of those who required assisted walking and 83.4% of those who required a wheelchair became bedridden. Conclusions Our data demonstrate that patients with fragility hip fractures have high morbidity and a mortality rate approaching 30%. Age and the ASA score significantly influence this high mortality rate.

Genetic Markers for Adolescent Idiopathic Scoliosis on Chromosome 19p13.3 among Saudi Arabian Girls.

STUDY DESIGN: Prospective case-controlled study. PURPOSE: This study aimed to assess genetic influence in Saudi Arabian children with adolescent idiopathic scoliosis (AIS). OVERVIEW OF LITERATURE: The genetic locus linked to chromosome 19p for idiopathic scoliosis has been described. A pilot study conducted at King Fahd Hospital of the University, Al-Khobar showed that three microsatellite markers (D19S216, D19S894, and DS1034) of chromosome 19p13.3 were significant in Saudi Arabian females compared with healthy subjects. METHODS: A total of 100 unrelated Saudi Arabian girls treated for AIS, their parents, healthy siblings, and healthy subjects were recruited for genetic analysis of markers on chromosome 19p13.3. After informed consent was obtained from their parents, blood samples were collected and parametric and nonparametric linkage analyses were performed using GENEHUNTER ver. 2.1. Multipoint linkage analysis was used to specify an autosomal dominant trait with a gene frequency of 0.01 and an estimated penetrance of 80% at the genotypic and allelic levels. RESULTS: Five hundred blood samples were collected and analyzed for microsatellite markers (D19S216, D19S894, and DS1034) of chromosome 19p13.3. Comparison among patients, family members, and healthy subjects revealed no significant association between markers and scoliosis at the genotypic level: D19S216 (p=0.21), D19S894 (p=0.37), and DS1034 (p=0.25). However, at the allelic level, a statistically significant association was observed for marker DS1034 (p=0.008), and marker D19S216 showed significance between fathers and patients (p<0.001) compared with patients and mothers. The other two markers, D19S216 (p=0.25) and D19S894 (p=0.17), showed no significant association between patients and mothers. CONCLUSIONS: At the allelic level, marker DS1034 was significantly associated with AIS patients and their fathers. This allelic marker on chromosome 19p13.3 appears to be important in AIS etiology.

Reassessment of osteoporosis-related femoral fractures and economic burden in Saudi Arabia.

UNLABELLED: The current study reassesses the prevalence of fragility fractures and lifetime costs in the Eastern Province of Saudi Arabia. Forty-two percent (391) of the fractures were at the neck of the femur, and 38.6 % (354) were inter-trochanteric fractures. The overall incidence was assessed to be 7528 (1,300,336 population 55 years or older) with the direct cost of SR564.75 million ($150.60 million). A National Fracture Registry and osteoporosis awareness programs are recommended. PURPOSE: Proximal femur fragility fractures are reported to be increasing worldwide due to increased life expectancy. The current study is carried out to assess the incidence of such fractures in the Eastern Province of Saudi Arabia and to assess the costs incurred in managing them annually. Finally, by extrapolating the data, the study can calculate the overall economic burden in Saudi Arabia. METHODS: The data of fragility proximal femur fractures was collected from 24 of 28 hospitals in the Eastern Province. The data included age, sex, mode of injury, type of fracture, prescribed drug (and its cost), and length of hospital stay. Population statistics were obtained from the Department of Statistics of the Saudi Arabian government Web site. RESULTS: Twenty-four hospitals (85 %) participated in the study. A total of 780 fractures were sustained by 681 patients. Length of stay in the hospital averaged 23.28 ± 13.08 days. The projected fracture rate from all the hospitals would be 917 (an incidence of 5.81/1000), with a total cost of SR68.77 million. Further extrapolation showed that the overall incidence could be 7528 (1,300,336 population 55 years or older) with the direct cost of SR564.75 million ($150.60 million). CONCLUSIONS: Osteoporosis-related femoral fractures in Saudi Arabia are significant causes of morbidity besides incurring economic burden. We believe that a National Fracture Registry needs to be established, and osteoporosis awareness programs should be instituted in every part of Saudi Arabia so that these patients can be diagnosed early and treated appropriately to reduce both the number of fractures and the economic burden of the fractures.

Effect of new neuronal growth factor on healing of sciatic nerve in rats.

OBJECTIVE: The study aimed to investigate the effect of a new peptide new nerve growth factor (NNGF) on the healing of divided sciatic nerves in rats. MATERIAL AND METHODS: Twenty Sprague-Dawley rats of 250-300g were divided into two groups (group 1 - study group and group 2 - control group). Under ketamine intramuscular anesthesia sciatic nerves were exposed, divided and repaired using 10/0 dexon. Study animals had 10mg/kg body weight of NNGF added to the repair. Electromyographic studies of the hind libs were carried out after 8weeks. The average stimulation was 50mA for 200µS and four twitches (T) were recorded. The animals were euthanized and the sciatic nerves were removed for histological analysis. RESULTS: There were no deaths in either of the groups. Electromyographic study showed that in the control group the average T1-T4 was 0.587±0.17% and in the study group the average was 87.89±5.02% (p value of 0.001). Histologically the control group showed regenerated axons sprouting from the proximal segment of cut nerve with empty endoneurial channels, while in the study group whole nerve trunks were seen within endoneurial channels. CONCLUSION: This study shows that the NNGF has a positive influence on the experimental healing of sciatic nerves in animals.

Healing of Experimentally Created Non-Union of Femur in Rats Using Bone Precursor Cells from Mesenchymal Stem Cells (MSCs).

BACKGROUND AND OBJECTIVE: Around 20% of fractures have impaired or no healing. Many procedures have been tried with varying success. The objective of this study is to assess effect of osteoblast transplant (obtained after proliferation and differentiation of MSCs of bone marrow aspirate) in healing of experimentally created non-union of femur in rats. METHODS: Non-Union of femur were created in Sprague-Dawley rats weighing 200-250 grams. In 20 rats, Femur fracture was surgically created in 20 rats and 2 mm of the periosteum was cauterized on each side of the fracture and this created a non-union in 8 weeks. In 10 animals bone marrow was aspirated from the femoral shaft using 24-gauge butterfly needle and injected in special media. The two groups 10 each were marked and animals were kept in the similar surroundings. After radiological confirmation of non-union at 8 weeks, an injection containing 1 x 10 (6) osteoblasts cells (1 million cells) dissolved in 200 microliters of balanced salt solution was injected at the nonunion site. In the control group of 10 rats 1 ml of normal saline was injected. In 5 animals of each group the fracture was fixed using 1 mm kirschner wire and the other 5 were treated without fixation. After 8 weeks of implantation the animals were radiographed and euthanized. The hind legs were disarticulated from the hip joints, specimens were stored in 2% formalin and histological evaluation was performed. RESULTS: There were no deaths in both the groups and there was one superficial infection in the control group. Eight weeks post implantation of the BM-MSCs derived osteoblasts, all the fractures of the study group united with robust mineralization and new bone formation confirmed by radiograph and histopathology. In the control group there was no healing and the histopathology showed full of fibrous tissue with cartilage cells lining the fracture site. CONCLUSIONS: In conclusion, our results indicate that implant of BM-MSCs derived osteo-progenitor cells at the non-union efficiently induces a complete union. We believe a similar study should be carried out in a larger animal before any human trials.

Topical delivery of vitamin d3: a randomized controlled pilot study.

BACKGROUND AND OBJECTIVE: The purpose of the present study is to explore the assessment if the transdermal delivery of vitamin D is feasible. METHODS: In 50 female Medical students, this study was conducted. Age, weight and height was taken, a detailed history and clinical examination was performed. Blood was drawn for 25 Hydroxy Vitamin D3 (25OHD) level. Two women had >30 ng/mL of 25OHD and was excluded from the study. The participants were divided into two groups of 24 in each arm. All participants equivocally agreed not to change their dietary habits and life style till the study was over. The study group of women were asked to apply; Top-D (Aloe Vera based- Vitamin D3) (Patency Pending) was developed at King Fahd Hospital of the University, AlKhobar with each gram of the Top-D cream delivering 5000 IU of vitamin D3. The second group used 1 gram of Aloe vera gel. The participants had no knowledge to which group they belong. A second blood sample was taken at the end of 3 months and the data was analyzed. RESULTS: The data of 48 women was available for analysis. The average age was 22.58 ± 1.95 years. The mean pre-treatment 25OHD in the study group was 12.05 ng/Ml ± 6.54 and post-treatment was 37.95 ng/mL ± 6.43 (P=0.001, CI<28.582 ). In control group pre-treatment 25OHD was 11.4 ng/mL ± 3.97 and post-treatment was 10.58ng/mL ± 3.03. CONCLUSIONS: This randomized control study shows that vitamin D3 can safely be delivered through the dermal route. This route could be exploited in treating vitamin D deficiency.

The effect of etoricoxib premedication on postoperative analgesia requirement in orthopedic and trauma patients.

OBJECTIVE: We hypothesized that etoricoxib premedication would reduce the need for additional opioids following orthopedic trauma surgery. METHODS: A double blind, controlled study, conducted in King Fahd University Hospital, King Faisal University, Dammam, Kingdom of Saudi Arabia. After obtaining the approval of the Research and Ethics Committee and written consent, 200 American Society of Anesthesiology grade I & II patients that underwent elective upper limb or lower limb fracture fixation surgeries during the period from August 2005 to October 2007 were studied. Patients were randomly premedicated using 120 mg of etoricoxib or placebo n=100, each. To alleviate postoperative pain, a patient controlled analgesia device was programmed to deliver one mg of morphine intravenously lockout time, 6 minutes. Visual analog scale and total postoperative morphine consumption over 24 hours and the adverse effects were recorded. RESULTS: One hundred patients in each group completed the study period. Etoricoxib premedication provides a statistically significant postoperative morphine sparing effect over 24 hours postoperatively. Total morphine consumption was 44.2 (8.2) in the placebo and 35.17.0mg in the etoricoxib groups p<0.001. The incidence of nausea and vomiting requiring treatment was lower in the etoricoxib group, p=0.014. The postoperative blood loss was similar in both groups. CONCLUSION: Etoricoxib is a suitable premedication before traumatic orthopedic surgery as it enhanced postoperative analgesia and reduced the need for morphine.

Are orthopedic surgeons prone to burnout?

OBJECTIVE: Burnout syndrome (BOS) is a state of physical, and emotional or mental exhaustion, depersonalization, and low personal accomplishment. Health care givers are most prone to suffer from BOS. There are no studies to date on BOS among trained orthopedic, and trauma surgeons. The objective of this study, was to assess the prevalence of BOS among the orthopedic surgeons in the Eastern province of Saudi Arabia. METHODS: This study was conducted among the orthopedic surgeons of the Eastern province of Saudi Arabia between September 2003 and October 2004. One hundred and two questionnaires of the Maslach Burnout Inventory (MBI) were sent to the qualified orthopedic surgeons with a self-addressed stamped envelope, from the Department of Orthopedic Surgery, King Fahd Hospital of the University, Al-Khobar. Three factors of MBI, which were assessed, were emotional exhaustion, depersonalization, and personal accomplishment. The data were entered in the database, and analyzed using Statistical Package for Social Science. RESULTS: Sixty-nine (67.6%) of the orthopedic surgeons completed the questionnaire. The average age was 45.72 +/- 6.82 (33-57) years. Thirty-five (50.7%) were found to be in a state of emotional exhaustion, 59.4% depersonalized, and 17% had low state of personal accomplishment. Doctors working in the government hospitals fared better than those in the private sector. CONCLUSION: Burnout syndrome is common among orthopedic surgeons working in the Eastern province Saudi Arabia. It is emphasized that awareness of the problem should be highlighted; programs need to be put in place to reduce the prevalence of burnout syndrome.