Callus Quality Research Articles

Induksi Kalus Krisan (Chrysanthemum morifolium var. reagen pink) dengan Penambahan Naphtalen Acetic Acid (NAA) dan Kinetin Secara In-Vitro

Chrysanthemum morifolium var. reagent pink is an ornamental flower that is widely cultivated in Indonesia. The increasing demand makes it difficult for farmers to fulfill the market. In-vitro tissue culture is an effective solution to multiply chrysanthemum seedlings in bulk and quickly. This study aims to identify the most effective concentration in inducing callus in chrysanthemum plants by combining Naphtalen Acetic Acid (NAA) and kinetin. This study was an experimental research with a completely randomized design (CRD), involving 12 treatments and each treatment was repeated 4 times. There were two treatment factors; NAA concentration (0 mg/l, 0.5 mg/l, 1 mg/l) and kinetin concentration (0 mg/l, 1 mg/l, 2 mg/l, 3 mg/l). Parameters in this study included days to callus appearance, percentage of callus explants, callus wet weight, callus color, and callus texture. Data were analyzed using analysis of variance (ANOVA). The results showed that (1) the fastest callus was formed at 11 HST by giving the treatment of 0 mg/l NAA + 3 mg/l kinetin; (2) the optimal treatment for the percentage of callus explants was obtained by giving 1 mg/l NAA + 1 mg/l kinetin); (3) the optimal treatment of callus wet weight was obtained by giving 1 mg/l NAA + 2 mg/l kinetin; (4) the best callus quality was obtained by giving a combination of NAA 1 mg/l and kinetin 1 mg/l, with a crumbly texture and brownish yellow callus.

Refeeding partially reverses impaired fracture callus in undernourished rats.

Adequate nutritional intake plays a crucial role in maximizing skeletal acquisition. The specific effects of a general food restriction and refeeding on fracture healing are yet to be fully understood. The aim of this study was to assess the impact of general food restriction and refeeding on fracture repair. Fifty-four male Wistar Hannover rats were randomly assigned into three groups: (1) Sham: Sham rats with femoral fracture; (2) FRes: Food-restricted rats with fracture, (3) Fres+Ref: Fres rats with refeeding. Following weaning, the FRes and Fres+Ref groups received 50% of the food amount provided to the Shams. In the sixth week of the experiment, all animals underwent a mid-right femur bone fracture, which was subsequently surgically stabilized. Following the fracture, the Fres+Ref group was refed, while the other groups maintained their pre-fracture diet. Bone calluses were analyzed on the fifth-day post-fracture by gene expression and on the sixth-week post-fracture using dual-energy X-ray absorptiometry, morphometry, histomorphometry, immunohistochemistry, computed microtomography, and torsion mechanical testing. Statistical significance was determined at a probability level of less than 0.05, and comparisons were made using the ANOVA test. Food restriction resulted in significant phenotypic changes in bone calluses when compared to sham rats characterized by deterioration in microstructure (i.e., BV, BV/TV, Tb.N, and Conn.D) reduced collagen deposition, bone mineral density, and mechanical strength (i.e., torque at failure, energy, and stiffness). Moreover, a higher rate of immature bone indicated a decrease in bone callus quality. Refeeding stimulated bone callus collagen formation, reduced local resorption, and effectively restored the microstructural (i.e., SMI, BCa.BV/TV, Tb.Sp, Tb.N, and Conn.D) and mechanical changes (i.e., torque at failure, energy, and angular displacement at failure) caused by food restriction. Despite these positive effects, the density of the bone callus, collagen deposition, and OPG expression remained lower when compared to the shams. Gene expression analysis didn't evidence any significant differences among the groups. Food restriction had detrimental effects on osseous healing, which was partially improved by refeeding. Based on these findings, new research can be developed to create targeted nutritional strategies to treat and improve fracture healing.

The Potential of Biotic Elicitors to Increase Isoflavone Production (Pachyrhizus erosus L.) Callus

Abstract The isoflavones contained in the Pachyrhizus plant (Pachyrhizus erosus Linn) have multiple functions, including serving as raw materials for the health/pharmacology or cosmetic industries. Isoflavones are beneficial as antioxidants, and cholesterol reducers and are capable of neutralizing free radicals and delaying aging. Tissue culture techniques are widely used to produce secondary metabolite compounds. Isoflavones can be made in vitro through callus production by tissue culture. By using elicitation techniques with the addition of various glucose concentrations to the culture media (media Murashige and Skoog or Vacin and Went), it is expected to increase the quantity and quality of callus and the content of secondary metabolites (Isoflavones) in Pachyrhizus callus. The research aims to increase the production of Isoflavones in Pachyrhizus callus by adding elicitors with various concentrations of glucose. The research method used a completely randomized factorial design. Factor I: two levels of basic media: M1 (Murashige and Skoog Media) and M2 (Vacin and Went Media). Factor II: Addition of Glucose concentrations with seven levels: 0%, 5%, 10%, 15%, 20%, 25%, 30%. The plant material used was the young leaves of Pachyrhizus. The results of the study showed that (1) all formed callus exhibited compact quality; (2) the addition of 30% glucose concentration resulted in the best growth of callus; (3) the highest Isoflavone content (0.15%) was found in the callus of Pachyrhizus erosus L. due to the treatment with both MS and VW media with 20% Glucose Elicitor

Biologic therapies in stress fractures: Current concepts

Stress fractures, a common overuse injury in physically active individuals, present a significant challenge for athletes and military personnel. Patients who sustain stress fractures have demanding training regimes where periods of rest and immobilisation have unacceptable negative consequences on sports goals and finances. Aside from being an overuse injury, there are various contributing risk factors that put certain individuals at risk of a stress fracture. The main two being nutritional deficiencies and hormonal variations, which have significant effects on bone metabolism and turnover. Historically, treatment of stress fractures focused on conservative strategies such as rest and immobilisation. Calcium and vitamin D deficiencies have been closely linked to stress fractures and so over time supplementation has also played a role in treatment. With the introduction of biologics into orthopaedics, newer treatment strategies have been applied to accelerate fracture healing and perhaps improve fracture callus quality. If such therapies can reduce time spent away from sport and activity, it would be ideal for treating stress fractures. This article aims to offer insights into the evolving landscape of stress fracture management. It investigates the pre-clinical evidence and available published clinical applications. Though fracture healing is well understood, the role of biologics for fracture healing is still indeterminate. Available literature for the use of biologic therapies in stress fractures are restricted and most reports have used biologics as a supplement to surgical fixation in subjects in studies that lack control groups. Randomised control trials have been proposed and registered by a few groups, with results awaited. Assessing individuals for risk factors, addressing hormonal imbalances and nutritional deficiencies seems like an effective approach to addressing the burden of stress fractures. We await better designed trials and studies to accurately determine the clinical benefit of adding biologics to the management of these injuries.

The value of ultrasonography in predicting the outcomes of simple long bone fractures treated by closed intramedullary nail fixation

PurposeUltrasonography has been used increasingly in orthopaedic practice credited to its low cost, easy accessibility, non-invasiveness, reproducibility, and safety from radiation. The purpose of this study was to test the validity and efficacy of ultrasonography as an adjunct in the assessment of fracture healing in long bones treated with intramedullary interlocking devices and its predictive value in determining the need for a secondary surgical procedure. MethodsThis was a descriptive longitudinal study of 40 skeletally mature patients who sustained long bone fractures of the tibia or femur treated using intramedullary interlocking nails. Patients with comminuted and segmental fracture patterns were excluded from the study. Each patient was evaluated at 6- and 12-week post-surgery using standard orthogonal radiographs and ultrasonography to assess fracture healing. Patients were then followed up until fracture union. Quantitative data was analyzed using frequency statistics and descriptive data with inferential statistics. ResultsUltrasonography predicted 87.5% union and 12.5% delayed or non-union as early as 6 weeks after surgery, while radiographs predicted 22.5% union as late as 3 months of follow-up. The sensitivity and specificity of ultrasonography in assessing fracture healing were 100% and 97.2%, respectively, with a positive predictive value of 80.0%. Vascular resistance index was less than 0.5 in all patients who developed delayed or non-union. ConclusionUltrasonography is able to predict fracture outcomes much earlier than standardized radiographs with comparable sensitivity and specificity. Vascular resistance index is an objective parameter in assessing callus quality and predicting fracture outcomes.

Plant Regeneration in Amorphophallus muelleri Blume. through Organogenic

Porang (Amorphophallus muelleri Blume.) usually used to functional food raw materials, caused its high glucomant content. Limitation of conventional propagation both vegetative and generative affected to decreasing of porang production to fulfil market demand. To solve the problem, plant tissue culture technique was chosen to rapid propagate bulbils of porang. So, the aim of this research to determine the combination and concentration of Plant growth Regulators (PGRs), such as 6-Benzylaminopurine (BAP), Naphthaleneacetic Acid (NAA), and Thidiazuron (TDZ) for quality of callus, also growth of callus and shoots for rapid propagation of porang, cause PGRs are one of factors that affect the success of in vitro propagation. Experimental design of this research was completely randomized designed with statistical analysis using analysis of variance (Anova) one way. Difference combination and concentration of cytokinin and auxin as manipulation variables, which several parameters growth of callus, shoots, and roots was observed to indicate the rapid organogenic of porang. The optimal shoot induction medium was Murashige and Skoog (MS) media with 5.0 mg.L-1 BAP and 0.2 mg.L-1 NAA on which number of shoots and shoot height reached 15±1.8 and (7.2±1.86) cm. After shoot formation stage, the proper media for rooting stage was determined, which was Murashige and Skoog (MS) media with 1.0 mg.L-1 NAA resulted the rooting rate reached 100% and roots appear earlier. Optimal combination and concentration of PGRs on micropropagation of Amorphophallus muelleri Blume. from bulbils explant should be known to increase the production of porang, which can fulfil the market demand.

IMPACT OF BMP-1 MODULATION IN EARLY FRACTURE REPAIR

BMP-1 is the major procollagen-C-peptidase activating, besides fibrillar collagen types I-III, several enzymes and growth factors involved in the generation of extracellular matrix. This study investigated the effect of adding and inhibiting BMP-1 directly post fracture.Standardised femoral fractures were stabilized by an intramedullary nail in 12 week-old female C57Bl/6J mice. We injected either 20 µL recombinant active BMP-1, activity buffer or the BMP-1 specific inhibitor “sizzled”. After 7, 14 and 28 days, mice were sacrificed. Femurs were dissected and paraffin slides were prepared. Callus composition was divided into soft tissue, mineralized and cartilaginous callus. Murine MC3T3 pre-osteoblastic cells were kept in culture adding BMP-1 and sizzled during osteoblastic differentiation. Putative cytotoxicity was determined using MTT-vitality assay. Cell calcification, collagen deposition, and BMP-2 and myostatin protein quantity were characterized.Adding BMP-1 displayed a weak positive effect on the outcome. After 7 days, more mineralised callus was present, meanwhile the cartilaginous callus was apparently remodelled at higher rate. In the case of BMP-1 inhibition, we observed more cartilaginous callus, which may indicate reduced stability. In cell culture, we could observe a high interference with mineralisation capabilities depending on the stage of osteoblastic development when adding BMP-1 or inhibiting it. Addition and inhibition impaired myostatin (anti-osteogen) and BMP-2 (pro-osteogen) expression.Interfering with BMP-1 homeostasis in this early stage of fracture repair seems to have rather negative effects. Inhibition apparently yields lower callus quality while the addition of BMP-1 does not significantly accelerate the healing outcome. Cell culture experiments show that BMP-1 application after 7 days of healing leads to higher collagen output but has no effect on mineralisation. This may suggest that BMP-1 application at a later time-point may lead to more pronounced beneficial effects on fracture repair.

Evaluation of grafting performances in Persian walnut affected by grafting methods and covering materials in Mugu district, Nepal

Walnut carries a great potential for cultivation in the high hills of Nepal, including the Mugu district. However, progress has not been as expected. Low graft survival rate and poor grafting practices account for inadequate number of quality saplings and consequently unreasonable price for successful ones. In this context, a field-based research was conducted in a two-factorial Randomized complete block design (RCBD) at Chhayanath Rara, 03, Mugu, to determine the suitable grafting method and covering material for grafting in walnut. Four different grafting methods viz. Tongue, saddle, cleft, and splice along with sawdust and grafting tape as graft covers were employed as treatments. The tongue grafting method yielded the highest callused rate (68.07%) and the highest graft take rate (48.07%). Similarly, sawdust used as a graft cover produced the most promising results on both the callused rate (56.37%) and the graft take rate (44.27%). Furthermore, the use of sawdust cover was found to have a greater callus quality (2.78) compared to that of using a grafting tape. No statistical differences were obtained in saleable plant rate and days to bud burst as a result of either of the factors. Apart from callus quality, the interaction between the two factors did not produce considerable variation in any of the variables. Hence, this study suggests the application of the tongue grafting method, along with the use of sawdust as a graft cover for enhanced grafting performance in agro-climatic zones similar to Mugu.

Is the association of electrochemotherapy and bone fixation rational in patients with bone metastasis?

Bone metastases are frequent in patients with cancer. Electrochemotherapy (ECT) is a minimally invasive treatment based on a high-voltage electric pulse combined with an anticancer drug. Preclinical and clinical studies supported the use of ECT in patients with metastatic bone disease, demonstrating that it does not damage the mineral structure of the bone and its regenerative capacity, and that is feasible and efficient for the treatment of bone metastases. Year 2014 saw the start of a registry of patients with bone metastases treated with ECT, whose data are recorded in a shared database. (1) Among patients who underwent ECT and internal fixation for bone metastasis, how many experienced a reduction of pain? (2) How many cases showed a radiological response? (3) How many patients presented local or systemic complication after ECT and fixation? Patients were treated in Bologna at Rizzoli Orthopaedic Institute between March 2014 and February 2022 and recorded in the REINBONE registry (a shared database protected by security passwords): clinical and radiological information, ECT session, adverse events, response, quality of life indicators, and duration of follow-up were registered. We consider only cases treated with ECT and intramedullary nail during the same surgical session. Patients included in the analysis were 32: 15 males and 17 females, mean age 65 ± 13 years (median 66, range 38-88 years), mean time since diagnosis of primary tumor 6.2 ± 7.0 years (median 2.9, range 0-22 years). Nail was indicated in 13 cases for a pathological fracture in, 19 for an impending fracture. Follow-up was available for 29 patients, as 2 patients were lost to follow-up and 1 was unable to return to controls. Mean follow-up time was 7.7 ± 6.5 months (median 5, range 1-24), and 16 patients (50%) had a follow-up longer than 6 months. A significant decrease in pain intensity was observed at the mean Visual Numeric Scale after treatment. Bone recovery was observed in 13 patients. The other 16 patients remained without changes, and one presented disease progression. One patient presented a fracture occurrence during the ECT procedure. Among all patients, bone recovery was observed in 13 patients: complete recovery in 1 patient (3%) and partial recovery in 12 patients (41%). The other 16 patients remained without changes, and one presented disease progression. One patient presented a fracture occurrence during the ECT procedure. However, healing was possible with normal fracture callus quality and healing time. No other local or systemic complications were observed. We found that pain levels decreased after treatment in 23 of the 29 cases for a pain relief rate of 79% at final follow-up. Pain is one of the most important indicators of quality of life in patients that undergo palliative treatments. Even if conventional external body radiotherapy is considered a noninvasive treatment, it presents a dose-dependent toxicity. ECT provides a chemical necrosis preserving osteogenic activity and structural integrity of bone trabeculae; this is a crucial difference with other local treatments and allows bone healing in case of pathological fracture. The risk of local progression in our patient population was small, and 44% experienced bone recovery while 53% of the cases remained unchanged. We observe intraoperative fracture in one case. This technique, in selected patients, improves outcome in bone metastatic patients combing both the efficacy of the ECT in the local control of the disease and the mechanical stability with the bone fixation to synergize their benefits. Moreover, the risk of complication is very low. Although encouraging data, comparative studies are required to quantify the real efficacy of the technique. Level of Evidence Level I, therapeutic study.

Is physeal distraction osteogenesis a good choice to treat CPT patients with tibial shortening?

The purpose of this study was to evaluate the healing index and complications of distraction osteogenesis through the physis in children with Congenital pseudarthrosis of the tibia (CPT) using Ilizarov external fixation device. Nine children with CPT who underwent distraction osteogenesis through the physis surgery with an Ilizarov external fixation device between June 2018 and August 2020 were included in the study. The average follow-up time was 36.1 months. X-ray examination was performed 1 month after physis lengthening, and the callus quality was evaluated using the Li classification. Any complications and the quality of the callus after tibial lengthening were followed up. The average age at the time of operation was 9.4 years, and the average follow-up time was 36.1 months. The average lengthening length of children with CPT was 6.6 cm. The incidence of good callus morphology was 100%, and the healing index was 51.1 d/cm. The average fixation time of the external fixation device was 316 days. Needle tract infection occurred in one case, ankle stiffness occurred in one case, ankle plantar flexion 20° deformity occurred in one case, angular deformity of the tibial lengthening segment occurred in one case (7°) and limited movement of knee joint occurred in one case (0-20°-70°). The osteogenic quality of the patients of distraction osteogenesis through the physis with tibial shortening and small diameter of the proximal tibia is good, which provides a choice of operation for the children with CPT.

In vitro propagation, shoot regeneration, callus induction, and suspension from lamina explants of Sorbus caloneura.

Sorbus caloneura has high ornamental and medicinal value but is endangered, and significant effort is required to preserve this natural resource. In this study, the stem and sterilized leaves of S. caloneura were used to explore the effects of different plant hormones basic medium type, initial callus quality, initial liquid volume, and ratio of old liquid culture medium to new on stem proliferation, regeneration and callus suspension culture. Naphthylacetic acid (NAA) and 6-benzyladenine (6-BA) significantly affected the proliferation of stem tissue. Murashige and Skoog (MS) basal medium supplemented with 1.75 mg/L 6-BA, 0.25 mg/L NAA, and 0.25 mg/L indole butyric acid (IBA) yielded a proliferation rate of 100%, with an average number of adventitious shoots per stem of 4.9. The best callus induction was observed with MS basal medium containing 0.5 mg/L 6-BA, 1 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D), and 0.2 mg/L thidiazuron (TDZ). The adventitious shoots were directly induced by MS basal medium containing 5.0 mg/L 6-BA, 0.5 mg/L NAA, and 1.5 mg/L kinetin (KT) and indirectly induced by medium containing 3.0 mg/L 6-BA, 0.3 mg/L NAA, and 0.1 mg/L TDZ. NAA significantly affected rooting rate, with ideal conditions found to be medium supplemented with 0.2 mg/L NAA and 1.5 mg/L IBA. MS basal medium with 4.5 g of calli and 120 mL of liquid culture medium without retention of the original culture medium yielded the best suspension effect for callus proliferation. Taken together, the results of this study lay a foundation for the breeding, preservation of germplasm resources, and genetic transformation of S. caloneura.

Risk-based Approach in Obtaining and Analysing the Characteristics of Baikal Skullcap Suspension Cultures

Introduction . The use of biologically active compounds of Baikal skullcap ( Scutellaria baicalensis Georgi) becomes especially important under pandemic conditions. The plant's value becomes the cause of overexploitation of the skullcap subterranean parts, which leads to the reduction of the population distribution range and inappropriate conditions of collection and harvesting. To prevent the extinction hazard of the species, in vitro cultivation of the plant's cells becomes a promising technology. This opens up the prospect of studying the physiology and biochemistry of plant cells, regulating the direction of valuable secondary metabolites synthesis and creating medicines without interfering with the natural habitat. Modern technologies provide opportunities to automate cell cultivation in order to minimize human involvement in the process. However, cultivation of plant cells is a probabilistic process that is influenced by a range of factors. The use of risk management tools will reduce the number of unintended consequences leading to dangerous and critical cell culture conditions. Aim . Risk analysis resulting from in vitro cultivation of Baikal skullcap ( Scutellaria baicalensis Georgi). Materials and methods . The object of the study was suspension cultures of Scutellaria baicalensis obtained from loose plant callus. Suspensions of Scutellaria baicalensis cells were grown in 250 ml flat-bottomed conical flasks under conditions of constant mixing on a shaker (rotation speed 100 rpm). The subculture cycle was 21 days. For microscopy of cell biomass, we used 0.1 % neutral red (NK) and 0,025 % Evans blue (EG) solution. Process risks were assessed using a cause-and-effect diagram (Ishikawa diagram). Results and discussion . It was found that the cells in the suspension cultures of skullcap were characterized by an elongated oval shape. Primary suspensions were distinguished by the presence of predominantly parenchymal cells (70 ± 2,0 %). A large aggregation was observed in the primary suspensions. The diameter of the aggregates reached about 8–15 mm. The growth properties of Baikal skullcap suspension depended on the density of the inoculum. As the amount of transplanted biomass decreased, cells showed a tendency to destruction. The results indicated the need for risk analysis of the laboratory technology of Baikal skullcap suspension cultivation. Five most important factors in plant cell cultivation were identified: callus quality, quality of prepared liquid nutrient medium, equipment serviceability and suitability, operator actions, and suitability of cell culture technology and biomass viability index sampling. Conclusion . An algorithm for analyzing the security of a Baikal skullcap suspension culture is proposed. The risk assessment can be used to make a conclusion about the probability of undesirable effects and their impact on the process. Risk management tools provide the basis for modernization of the available technology and automatic outfitting of the process.

Vibration therapy as an effective approach to improve bone healing in diabetic rats.

ObjectiveTo investigate the effects of vibration therapy on fracture healing in diabetic and non-diabetic rats.Methods148 rats underwent fracture surgery and were assigned to four groups: (1) SHAM: weight-matched non-diabetic rats, (2) SHAM+VT: non-diabetic rats treated with vibration therapy (VT), (3) DM: diabetic rats, and (4) DM+VT: diabetic rats treated with VT. Thirty days after diabetes induction with streptozotocin, animals underwent bone fracture, followed by surgical stabilization. Three days after bone fracture, rats began VT. Bone healing was assessed on days 14 and 28 post-fracture by serum bone marker analysis, and femurs collected for dual-energy X-ray absorptiometry, micro-computed tomography, histology, and gene expression.ResultsOur results are based on 88 animals. Diabetes led to a dramatic impairment of bone healing as demonstrated by a 17% reduction in bone mineral density and decreases in formation-related microstructural parameters compared to non-diabetic control rats (81% reduction in bone callus volume, 69% reduction in woven bone fraction, 39% reduction in trabecular thickness, and 45% in trabecular number). These changes were accompanied by a significant decrease in the expression of osteoblast-related genes (Runx2, Col1a1, Osx), as well as a 92% reduction in serum insulin-like growth factor I (IGF-1) levels. On the other hand, resorption-related parameters were increased in diabetic rats, including a 20% increase in the callus porosity, a 33% increase in trabecular separation, and a 318% increase in serum C terminal telopeptide of type 1 collagen levels. VT augmented osteogenic and chondrogenic cell proliferation at the fracture callus in diabetic rats; increased circulating IGF-1 by 668%, callus volume by 52%, callus bone mineral content by 90%, and callus area by 72%; and was associated with a 19% reduction in circulating receptor activator of nuclear factor kappa beta ligand (RANK-L).ConclusionsDiabetes had detrimental effects on bone healing. Vibration therapy was effective at counteracting the significant disruption in bone repair induced by diabetes, but did not improve fracture healing in non-diabetic control rats. The mechanical stimulus not only improved bone callus quality and quantity, but also partially restored the serum levels of IGF-1 and RANK-L, inducing bone formation and mineralization, thus creating conditions for adequate fracture repair in diabetic rats.

Outcomes of two circular external fixation systems in the definitive treatment of acute tibial fracture related infections

IntroductionAcute tibial fracture-related infection (FRI) is one of the most feared and challenging complications after a tibial fracture. The most appropriate treatment in this scenario is far from a resolved topic. Circular external fixators (CEFs) offer multiplanar control and minimize soft tissue injury using temporary implants far from the infected area. This study aimed to investigate the outcomes of two different types of CEFs (Ilizarov and hexapod) in the treatment of a series of acute tibial FRIs. Material and MethodsA retrospective study at two specialized limb reconstruction centres identified all patients with an acute tibial FRI (≤4 weeks after index procedure) definitively treated with a CEF from January 2015 to December 2020. Primary outcomes: fracture healing and infection eradication rate with a minimum FU of 12 months after frame removal. Secondary outcomes: to investigate the differences between the two types of circular frames regarding final post-treatment deformity magnitude. ResultsWe included 31 patients with acute tibial FRIs: 18 treated with hexapod-type and 13 with Ilizarov-type CEFs. Average age was 45.5±16.56 years. Fracture healing and infection eradication were achieved in all patients (31/31) after a mean follow-up of 24.7 months (range 12.1-55.3). Patients treated with an Ilizarov-type fixator presented shorter time to fracture union (5.5±2.2 months vs. 9.2±6.0 months; p-value 0.021) and shorter duration of external fixation (p-value 0.001). Regarding residual post-treatment deformity, the hexapod system presented significantly less residual coronal translation deformity (p-value 0.034) and better callus quality. Fixator-related complications were similar when comparing the two groups. No significant differences were seen in pain (p-value 0.25), RTW rate (35% vs. 45%; p-value 0.7) or functionality (p-value 0.4). ConclusionsDefinitive circular external fixation is an excellent treatment for acute tibial FRI. Both Ilizarov and hexapod systems offer a very high rate of fracture healing and infection eradication. Although both presented very low radiological post-operative residual deformity, the hexapod system showed less residual coronal translation deformity and better callus quality.

The Relationship Between Limb Lengthening Rate and Callus Quality in Patients with Achondroplasia.

This study investigated the effect of the femur, tibia, and humeral lengthening rate in patients with achondroplasia and regenerated bone quality. The records of the patients with achondroplasia who underwent limb lengthening surgery for both upper and lower extremities between 2002 and 2019 were retrospectively reviewed. Bone formation regeneration was evaluated in each segment at anteroposterior and lateral radiographs and the callus quality was determined at the first month of the consolidation period according to Li's classification system. This study included 42 (28 females and 14 males), 38 (26 females and 12 males), and 17 (11 females and 6 males) patients with bilateral femoral, bilateral tibial, and bilateral humeral lengthening. The mean lengthening rate was 0.920 ± 0.23 (range, 0.53-1.67), 0.813 ± 0.17 (range, 0.51-1.26), and 1.02 ± 0.26 (range, 0.58-150) mm/day in the femoral, tibial, humeral groups, respectively. In the femoral group, 75% femur with good morphological quality, 56.6% good morphological quality in tibial group and 55.9% good morphological quality in humeral group. Statistically significant relationships were found between femoral lengthening rate and callus quality (p < 0.001; r = 0.454). However, no significant correlation was found in the humeral and tibial groups. Moreover, the sensitivity and specificity of the lengthening rate for obtaining good morphological quality callus were 72% and 80%, respectively, with an optimum diagnostic cutoff value of 0.976mm/day for femoral lengthening. A higher-rate good morphological callus was obtained in femoral lengthening compared with tibia and humerus in patients with achondroplasia.

Three-dimensional printable metamaterial intramedullary nails with tunable strain for the treatment of long bone fractures

Novel metamaterial unit cells with tunable strain were incorporated into titanium intramedullary (IM) nails to overcome problems of dense metallic implants. Three-dimensional (3D) models of cylindrical, pillar, and reference metamaterials were tuned and analyzed for structural authentication. Further, one solid IM nail (S0) and four different cylindrical metamaterial cell IM nails with gap sizes of 0.15 mm, 0.30 mm, 0.45 mm, and 0.60 mm were used to achieve 5% (S1), 10% (S2), 15% (S3), and 20% (S4) strain, respectively, in a 3-mm fracture, to investigate the healing outcomes. The bending stiffness (callus quality) in the S2 metamaterial IM nail increased by 22% between the 7th and 16th iterations, whereas for the S0, S1, S3, and S4 IM nails, the increases were 5.6%, 8.1%, 12.4%, and 9.0%, respectively. The S2 IM nail exhibited the best healing and bending stiffness; however, the S0 IM nail exhibited the worst results. This unique design was customized for different fracture gap sizes according to patient-specific needs.

The Effect of Diclofenac Sodium on Callus Formation in White Male Rat (Rattus norvegicus) Cruris Fracture Healing

Highlights: Sodium diclofenac is one of NSAID a common treatment to relieve pain associated with bone fractures. Sodium diclofenac with a some dose of body weight could decrease the callus quality on fracture healing. Abstract: Non-steroidal anti-inflammatory drugs (NSAIDs), such as diclofenac sodium, are standard treatments to relieve pain associated with bone fractures. The bone healing process consists of four stages: inflammation, soft callus formation, complex callus formation, and bone remodeling. Previous studies mentioned that intake of NSAIDs (sodium diclofenac) could inhibit the bone healing process. This study examined the effect of diclofenac sodium intake on callus formation in fracture healing. In this study, thirty-six rats (Rattus Norvegicus) with fractures were used and divided into two groups, namely 18 rats for the control and 18 rats for the treatment group. In the treatment group, each rat was given 1.8 mg sodium diclofenac/150 grams of body weight per day. In the control group, each rat was given CMC-Na 0.5% with equal volume as diclofenac sodium in the treatment group. After 28 days, all the rats were stunned until dead, and the diameter and strength of their calluses were measured. In the treatment group with diclofenac sodium1.8 mg/ 150 grams BW/ 28 days after the tibia bone callus was pressed using the Shimadzu tool, the lowest callus strength was found to be 56.500 N, and the highest callus strength was 59.000 N. The lowest callus diameter in the treatment group was 4 mm, the highest was 5 mm. In the control group, the lowest callus strength was 76 N, and the highest callus strength was 77 N. The lowest callus diameter in the control group was 6 mm, and the highest was 8 mm. The strongest callus in the treatment group was found in the sixth observation, with a value of 59 N and a diameter of 4 mm. In the control group, the highest callus strength was 77 N, with a diameter of 7-8 mm. These measurements were found on the 5th, 7th, 8th, 9th, 16th, and 17th observations. Diclofenac sodium with a dose of 1.8 mg/150 grams of body weight could decrease the callus quality parameters, such as callus strength and diameter on fracture healing.

Fracture-related infection in osteoporotic bone causes more severe infection and further delays healing.

AimsWith the ageing population, fragility fractures have become one of the most common conditions. The objective of this study was to investigate whether microbiological outcomes and fracture-healing in osteoporotic bone is worse than normal bone with fracture-related infection (FRI).MethodsA total of 120 six-month-old Sprague-Dawley (SD) rats were randomized to six groups: Sham, sham + infection (Sham-Inf), sham with infection + antibiotics (Sham-Inf-A), ovariectomized (OVX), OVX + infection (OVX-Inf), and OVX + infection + antibiotics (OVX-Inf-A). Open femoral diaphysis fractures with Kirschner wire fixation were performed. Staphylococcus aureus at 4 × 104 colony-forming units (CFU)/ml was inoculated. Rats were euthanized at four and eight weeks post-surgery. Radiography, micro-CT, haematoxylin-eosin, mechanical testing, immunohistochemistry (IHC), gram staining, agar plating, crystal violet staining, and scanning electron microscopy were performed.ResultsAgar plating analysis revealed a higher bacterial load in bone (p = 0.002), and gram staining showed higher cortical bone colonization (p = 0.039) in OVX-Inf compared to Sham-Inf. OVX-Inf showed significantly increased callus area (p = 0.013), but decreased high-density bone volume (p = 0.023) compared to Sham-Inf. IHC staining showed a significantly increased expression of TNF-α in OVX-Inf compared to OVX (p = 0.049). Significantly reduced bacterial load on bone (p = 0.001), enhanced ultimate load (p = 0.001), and energy to failure were observed in Sham-Inf-A compared to Sham-Inf (p = 0.028), but not in OVX-Inf-A compared to OVX-Inf.ConclusionIn osteoporotic bone with FRI, infection was more severe with more bone lysis and higher bacterial load, and fracture-healing was further delayed. Systemic antibiotics significantly reduced bacterial load and enhanced callus quality and strength in normal bone with FRI, but not in osteoporotic bone.Cite this article: Bone Joint Res 2022;11(2):49–60.

Effect of Different Factors on Callus Induction from Leaf Explants of &lt;i&gt;Jatropha curcas&lt;/i&gt; L.

In order to further optimize the tissue culture regeneration system of Jatropha curcas, the leaf explants of J. curcas were used as experimental materials, and MS medium supplemented with 0.5 mg/L 6-BA, 0.1 mg/L TDZ and 0.5 mg/L IBA was used as the basic medium. The effects of callus induction from leaf explants of J. curcas was studied by added different concentrations of compound sodium nitrophenolate (CSN), silver nitrate and diethyl aminoethyl hexanoate (DA-6) into the basic medium. The results showed that adding 8 mg/L CSN or 10 mg/L DA-6 in the basal medium could increase the callus induction rate of leaf explants. While the number of callus induced by basal medium supplemented with silver nitrate was decreased, but the quality of callus was improved.

In vitro Study: The Potential for Papain Production from Papaya Leaf Callus

Normally papaya plant can create sap white when tapped or in hur. Sap white it very well may be created on the entirety of its parts like natural product, blossoms and leaves petiole. Papain is a proteolytic catalyst that is created by secluding papaya sap. Papain is valuable as a breakdown of protein particles, and is regularly utilized by numerous individuals in different kinds of mechanical fields. The advantage of papain are so assorted thatb it has been broadly need for improvement perposes in industry, medication and creature feed. Papain creation can be developmentt with Biotechnology tissue culture .With improving the creation of callus plantled outgrowth leaves can expanded the cantaint of papain. The exploration goals were to decide the sorts of papaya assortments whose leaf callus could create the best papain. Realizing the papain content that can be created from three assortments of papaya leaves planted on MS and VW media. The exploration strategy utilized a totally randomized plan. Factor I was papaya assortments with 3 levels (California, Bangkok, and Gantung ). The subsequent factor is the sort of culture media (MS and VW). There are 6 mixes. Every treatment mix is rehashed multiple times with 10 test tests. The outcomes appeared; (1) There was a cooperation between the treatment of the concentrate source from the leaves of 3 sorts of Papaya assortments and the utilization of MS and VW media on callus amount and callus quality boundaries. (2 The amount and nature of callus on MS media was better compared to in VW media. (3) Callus of leaves California assortment produce Papain best than Bangkok and Gantung Variety. (3) Research result show that : all callus produces papain. The callus papaya all assortment containing papain range 11,86 % - 19,50 %.. The best found in treatment papain M1V1 ( MS Media - California (19,50 %); M2V1 (VW Media – California) 18,06 % papain content