Autologous adipose-derived mesenchymal stem cells and hydroxyapatite for bone defect in rabbits

This study aims to evaluate the effect of autologous adipose-derived mesenchymal stem cells (AAD-MSC), with and without synthetic absorbable hydroxyapatite (HAP-91), on the bone regeneration in rabbits. Thirty-four female white New Zealand rabbits were submitted to a 10 mm distal diaphyseal radius ostectomy, divided into 3 experimental groups according to the treatment established. The bone gap was filled with 0.15 ml of a 0.9% saline solution containing two million AAD-MSC (G1), or AAD-MSC associated with HAP-91 (G2). The control group (CG) received only 0.15 ml of the 0.9% saline solution. Radiographs were made post-operatively, and after 15, 30, 45 and 90 days. Fifty percent of the samples were submitted to a histological examination at 45 days and the remaining ones at 90 days post-operatively. Radiographically, the periosteal reaction, bone callus volume and bone bridge quality were superior in G2 (P < 0.05). Histologically, the bone repair was faster and more efficient in G1 at 45 days (P < 0.05). In conclusion, AAD-MSC improved the regeneration on the experimentally induced bone defects in rabbits; however, the use of hydroxyapatite requires caution given the granulomatous reaction produced in the species.

Systemic Administration of Recombinant Irisin Accelerates Fracture Healing in Mice

To date, pharmacological strategies designed to accelerate bone fracture healing are lacking. We subjected 8-week-old C57BL/6 male mice to closed, transverse, mid-diaphyseal tibial fractures and treated them with intraperitoneal injection of a vehicle or r-irisin (100 µg/kg/weekly) immediately following fracture for 10 days or 28 days. Histological analysis of the cartilaginous callus at 10 days showed a threefold increase in Collagen Type X (p = 0.0012) and a reduced content of proteoglycans (40%; p = 0.0018). Osteoclast count within the callus showed a 2.4-fold increase compared with untreated mice (p = 0.026), indicating a more advanced stage of endochondral ossification of the callus during the early stage of fracture repair. Further evidence that irisin induced the transition of cartilage callus into bony callus was provided by a twofold reduction in the expression of SOX9 (p = 0.0058) and a 2.2-fold increase in RUNX2 (p = 0.0137). Twenty-eight days post-fracture, microCT analyses showed that total callus volume and bone volume were increased by 68% (p = 0.0003) and 67% (p = 0.0093), respectively, and bone mineral content was 74% higher (p = 0.0012) in irisin-treated mice than in controls. Our findings suggest that irisin promotes bone formation in the bony callus and accelerates the fracture repair process, suggesting a possible use as a novel pharmacologic modulator of fracture healing.

Effect of salinity and drought stress on morphological and biochemical properties of two Iranian fenugreek (Trigonella foenum-graecum) populations

In this study, micro propagation of two Iranian fenugreek populations and their morphological and biochemical responses to salinity and drought stresses in in vitro culture condition were conducted using factorial experiment in a completely randomized design in three replications. Different explant type (terminal bud, cotyledon and epicotyledon explant) were cultured in MS medium contain different concentration of plant growth regulators such as kin (0, 0.5 and 1 mg / l) and 2,4-D (0.5, 1 and 2 mg / l). Murashige and Skoog (MS) medium supplemented with 1 mg/l kinetin and 2 mg/l 2,4-D showed the highest callus proliferation rate per explants in both populations. The highest amount of callus volume was obtained from the explants of the terminal bud. Proliferated calli from terminal bud explant were green and yellowish, from cotyledon were yellowish to white with soft texture, and the cotyledons were greenish and compact. The results of salinity stresses with different concentrations of sodium chloride (0, 70 and 120 mM) and drought stress with polyethylene glycol (0, 5 and 10%) showed that both stresses decreased callus growth and increased total protein, proline, catalase, peroxidase and trigonelin content in both populations. Trigonelin measurement showed that ‘Borazjan’ papulation had higher trigonelin content, in vitro, than ‘Ardestan’ papulation.

Repurposing denosumab for recalcitrant bone healing

Fracture healing has four phases: haematoma formation, soft callus, hard callus and remodelling. Often, non-healing fractures have an arrest of one of these phases, which need resurgery. We have repurposed denosumab for impaired fracture healing cases to avoid surgical intervention. Here, we report a series of three cases of impaired fracture healing where denosumab was given 120 mg subcutaneous dosages for 3 months to enhance healing. All the three cases have shown complete bone union at a mean follow-up of 6.7 months (5–9 months) as assessed clinically and radiologically, and have observed no adverse effect of the therapy. Denosumab given in this dose aids fracture healing by increasing callus volume, density and bridges the fracture gap in recalcitrant fracture healing cases where the callus fails to consolidate.

Prognostic Significance of Alkaline Phosphatase Activity and Callus Volume in Operatively Treated Bone Fractures

Total and bone alkaline phosphatase are indicators of bone formation, a process essential in bone healing. The aim of this study was to assess the relationship of both total and bone alkaline phosphatase with the course and features of healing in surgically treated long bone fractures as compared to the callus volume. In this study, total and bone alkaline phosphatase levels and the callus volume were measured in two patients with long bone fractures. Fracture healing was rapid in one patient and slow in the other. Depending on the healing outcome, on day 7 an increase in the case of slow healing and a decrease in the case of rapid healing was noted for both total and bone alkaline phosphatase. In the case of slow healing, the callus volume was significant whereas in the case of rapid healing the callus was almost invisible on day 7. This result indicates a possible prognostic significance of aforementioned clinical biochemical and radiographic parameters in the monitoring of long bone fracture healing.

EMBRIOGENESIS SOMATIK DARI KALUS MANGGIS (Garcinia mangostana L.) ASAL BENGKALIS DENGAN PEMBERIAN BAP DAN MADU SECARA IN VITRO

AbstrakGarcinia mangostana L. dikenal dengan sebutan queen of the tropical fruits. Buah manggis terbentuk secara apomiksis yang bersifat rekalsitran. Salah satu cara perbanyakan tanaman manggis adalah dengan teknik kultur in vitro melalui embriogenesis somatik. Embriogenesis somatik manggis dilakukan dengan pembentukan kalus terlebih dahulu. Penelitian ini bertujuan untuk menentukan konsentrasi terbaik BAP dan madu secara tunggal serta kombinasinya dalam pembentukan embriogenesis somatik pada kalus biji manggis asal Bengkalis. Penelitian ini menggunakan rancangan acak kelompok (RAK) dengan pemberian konsentrasi BAP (3 dan 7 mg/L) dan madu (3, 6, dan 9 mL/L), secara baik tunggal maupun kombinasi, pada media Murashige-Skoog (MS) dengan 3 ulangan. Hasil penelitian menunjukkan bahwa pemberian BAP dan madu dalam seluruh perlakuan tersebut berpengaruh terhadap pembentukan fase-fase embriogenesis somatik kalus manggis. Konsentrasi terbaik dalam pembentukan fase embriogenesis somatik diperoleh dari perlakuan 3 mg/L BAP + 9 mL/L madu dengan presentase pembentukan kalus 100%, waktu muncul kalus 10,67 hst, volume kalus 1,33 dan adanya fase embriogenesis somatik berupa globular, hati, dan torpedo.Abstract Garcinia mangostana L. was known as the queen of the tropical fruits. Mangosteen was formed by apomixis which is recalcitrant. One of the methods of mangosteen propagation is by using a tissue culture technique through somatic embryogenesis. Mangosteen somatic embryogenesis occurs preceded by callus formation. This study aimed to determine the best concentration of BAP and honey in single as well as in combination for the formation phase of somatic embryogenesis in the callus of mangosteen from Bengkalis. The study used a randomized block design with the addition of BAP (3 and 7 mg/L) and honey (3; 6; and 9 mL/L) either single or combination in Murashige-Skoog (MS) medium with 3 replications. The results of this study indicated that the addition of BAP and honey in all treatments affected the phases of somatic embryogenesis of mangosteen callus. The best concentration in the formation of somatic embryogenesis was obtained from, the treatment of 3 mg/L BAP + 9 mL/L which produced 100% of callus formation, with callus emergence time of 10.67 days after plantation, callus volume of 1.33 and the presence of somatic embryogenesis in the form of globular, heart, and torpedo.

Basic Fibroblast Growth Factor-Anchored Multilayered Mesenchymal Cell Sheets Accelerate Periosteal Bone Formation

Cell-based regenerative therapy has the potential to repair bone injuries or large defects that are recalcitrant to conventional treatment methods, including drugs and surgery. Here, we developed a multilayered cell-based bone formation system using cells coated with fibronectin-gelatin (FN-G) nanofilms. The multilayered mesenchymal cells (MLMCs) were formed after two days of culture and were shown to express higher levels of BMP-2 and VEGF compared to monolayer cultures of MCs. The MLMCs were used as a graft material in combination with a fusion protein consisting of basic fibroblast growth factor (bFGF), polycystic kidney disease (PKD) domain, and the collagen-binding domain (CBD) of Clostridium histolyticum collagenase. In femur sites grafted with the MLMCs, significantly higher levels of callus volume and bone mineral content were observed compared to the sham controls. The callus volume and bone mineral content were further increased in femur sites grafted with bFGF-PKD-CBD/MLMCs. Taken together, these results suggest that bFGF-PKD-CBD/MLMCs, which can be simply and rapidly generated in vitro, have the potential to promote bone repair when grafted into large defect sites.

Improved Fracture Healing in Patients with Concomitant Traumatic Brain Injury: Proven or Not?

Over the last 3 decades, scientific evidence advocates an association between traumatic brain injury (TBI) and accelerated fracture healing. Multiple clinical and preclinical studies have shown an enhanced callus formation and an increased callus volume in patients, respectively, rats with concomitant TBI. Over time, different substances (cytokines, hormones, etc.) were in focus to elucidate the relationship between TBI and fracture healing. Until now, the mechanism behind this relationship is not fully clarified and a consensus on which substance plays the key role could not be attained in the literature. In this review, we will give an overview of current concepts and opinions on this topic published in the last decade and both clinical and pathophysiological theories will be discussed.