Evaluation of bone marrow-derived mesenchymal stem cells with eggshell membrane for full-thickness wound healing in a rabbit model.

Biomaterials capable of managing wounds should have essential features like providing a natural microenvironment for wound healing and as support material for stimulating tissue growth. Eggshell membrane (ESM) is a highly produced global waste due to increased egg consumption. The unique and fascinating properties of ESM allow their potential application in tissue regeneration. The wound healing capacity of bone marrow-derived mesenchymal stem cells (BM-MSCs), ESM, and their combination in rabbits with full-thickness skin defect (2 × 2 cm2) was evaluated. Twenty-five clinically healthy New Zealand White rabbits were divided into five groups of five animals each, with group A receiving no treatment (control group), group B receiving only fibrin glue (FG), group C receiving FG and ESM as a dressing, group D receiving FG and BM-MSCs, and group E receiving a combination of FG, ESM, and BM-MSCs. Wound healing was assessed using clinical, macroscopical, photographic, histological, histochemical, hematological, and biochemical analysis. Macroscopic examination of wounds revealed that healing was exceptional in group E, followed by groups D and C, compared to the control group. Histopathological findings revealed improved quality and a faster rate of healing in group E compared to groups A and B. In addition, healing in group B treated with topical FG alone was nearly identical to that in control group A. However, groups C and D showed improved and faster recovery than control groups A and B. The macroscopic, photographic, histological, and histochemical evaluations revealed that the combined use of BM-MSCs, ESM, and FG had superior and faster healing than the other groups.

Selection and validation of suitable reference gene for qPCR gene expression analysis in lamb testis cells under Sheep pox virus infection.

Virus infection alters host gene expression, therefore ideal and stable reference housekeeping genes are required to normalise the expression of other expressed host genes in quantitative real-time PCR (qRT-PCR). The suitable reference gene may vary in response to different viral infections in different hosts or cells. In the present study, we cultured primary lamb testis cells (LTC) and assessed the expression stability of seven widely used housekeeping genes (B2M, HMBS, HPRT1, HSP-90, POLR2A, 18s_RNA, GAPDH) as reference genes in Sheeppox virus (SPPV) infected and control (uninfected-0h) LTC at 0.5h, 4.0h, 8.0h, and 12.0h post-infection) using NormFinder, Bestkeeper, geNorm, and the comparative ΔCT method in RefFinder based on their expression levels. Analysis revealed that HSP90, 18s_RNA, HPRT, POLR2A, and B2M were the most stable genes from the panel in the individual analysis group in 0h, 0.5h, 4.0h, 8.0h, and 12.0h, respectively. Furthermore, B2M was shown to be the most stable reference gene in the combined control with the respective and overall infected groups, except the control group of 4.0hpi of SPPV infection. In this study, we selected the most suitable reference genes in LTC for particular time points of SPPV infection. The identified most suitable housekeeping gene can be used during normalization of expression of other targeted genes at aspecific time point of SPPV infection.

Development of a novel atrophic non-union model in rabbits: A preliminary study.

BACKGROUND AND AIM: The currently available atrophic non-union models rely on wide segmental excision of bone diaphysis to impede the process of healing but lack resemblance to the clinical scenario. The present study focused on developing an in vivo model of atrophic non-union fracture in rabbit radius that can replicate the clinical scenario. MATERIALS AND METHODS: The atrophic non-union fracture model was developed by creating a 10 mm segmental bone defect in the radial diaphysis of five adult New Zealand White rabbits. The periosteum (2 mm) of the cut bone ends was cauterized using electrocautery to induce atrophy. Atrophic non-union was confirmed using radiographic and histologic evaluations on 30th postoperative day. RESULTS: The radiographic signs of healing were completely absent in all the rabbits on 30th postoperative day, indicating inert bone ends. Histological findings further confirmed the presence of inert bone ends, indicating the development of atrophic non-union. CONCLUSION: The combination of the segmental bone defect, electrocautery induced thermal damage of bone end periosteum, and delayed treatment can induce the development of atrophic non-union fracture model in rabbits that can replicate the clinical scenario.

Classification and coding of platelet-rich plasma derived from New Zealand white rabbits for tissue engineering and regenerative medicine applications.

BACKGROUND AND OBJECTIVE: Platelet-rich plasma (PRP) is a category of platelet concentrate that has been widely used as a therapeutic modality in musculoskeletal medicine. The present study was conducted to classify and code the non-activated platelet-rich plasma (nPRP) derived from New Zealand white rabbits for tissue engineering and other regenerative medicine applications. METHODS: PRP was prepared from the whole blood by double centrifugation protocol using a laboratory centrifuge. The prepared nPRP was characterized using the parameters such as platelet dose, the relative composition of platelets, WBC, and RBC. The production protocol was described using the parameters such as platelet enrichment factor, factor increase in WBC concentration, platelet capture efficiency, WBC-reducing efficiency, and RBC-reducing efficiency. The nPRP was also classified and coded using the most recent and universally accepted classification and coding systems. RESULTS: The non-activated leukocyte-poor red cell-rich PRP had an average platelet count of 1875.75 × 109/L, which is higher than the basal platelet concentration in the whole blood. Furthermore, the protocol used in this study has a mean platelet capture efficiency of 47.43 ± 6.42%. CONCLUSION: The protocol described in this study can be used to prepare non-activated leukocyte-poor red cell-rich PRP (Red-PRP IC1) from rabbits that can be coded as 318-00-00.

Standardization and characterization of adipose-derived stromal vascular fraction from New Zealand white rabbits for bone tissue engineering.

BACKGROUND AND AIM: Adipose tissue-derived stromal vascular fraction (SVF) contains a heterogeneous cell population comprising multipotent adipose-derived stem cells. Regenerative therapy using adipose-derived SVF has broad applications in bone tissue engineering due to the superior osteogenic potential of SVF. This study was designed to standardize and characterize adipose-derived SVF obtained from New Zealand white rabbits for bone tissue engineering and other potential applications. MATERIALS AND METHODS: Ten skeletally mature and clinically healthy adult New Zealand white rabbits were used in this study. The SVF was prepared using surgically resected interscapular adipose tissue following enzymatic digestion with 0.1% collagenase type I solution. The SVF pellet obtained after the final centrifugation step was suspended in a 0.5 mL control solution to obtain ready-to-use adipose-derived SVF. The freshly prepared SVF was characterized based on the total SVF cell count and cell yield per gram of adipose tissue. The SVF cells were enumerated using a hemocytometer. RESULTS: Interscapular adipose tissue depots are ideal for preparing autologous adipose-derived SVF due to the ease of access. The interscapular adipose-derived SVF prepared by enzymatic digestion had an average cell yield of 3.15±0.09×106 cells/g adipose tissue. Freshly prepared SVF had a total cell count of 3.15±0.09×104 cells/µL. CONCLUSION: The enzymatic digestion of adipose tissue using 0.1% collagenase resulted in better cell yield per gram than methods previously reported in rabbits. The use of adipose-derived SVF can preclude the requirement for an additional culture period. In addition, it may also reduce the risk of extensive cell contamination, which makes it a safe and cost-effective strategy for bone tissue engineering applications.

Diagnosis and surgical management of an intraocular foreign body secondary to ballistic wound in a Rhesus macaque (Macaca mulatta).

BACKGROUND: Intraocular foreign bodies (IOFBs) such as air gun pellet is a rare finding in wild animals like Rhesus macaque (Macaca mulatta). The purpose of the present scientific report is to describe the surgical retrieval of IOFB secondary to ballistic wound in a wild Rhesus macaque. CASE DESCRIPTION: A juvenile female wild Rhesus macaque was brought with the history of swollen and inflamed right eye for the past several days. FINDINGS/TREATMENT AND OUTCOME: Clinical examination revealed presence of partially healed wound over the dorsal eyelid. Radiographic examination revealed the presence of a metallic foreign body inside the right orbit. Inflamed and persistently closed eyelid prevented the further localization of the metallic foreign body. Lateral canthotomy was performed under general anesthesia. Following the failure to recover the metallic foreign body from the ocular adnexa, right eye vitrectomy was performed to retrieve the IOFB. The metallic foreign body was recovered from the posterior chamber of the right eye. Due to the poor prognosis of the already damaged eye, enucleation of the eye was performed which was followed by tarsorrhaphy. Further examination of the foreign body identified it as a 4.5 mm (.177 Calibre) air gun pellet. Post-operatively animal was treated with antibiotics and anti-inflammatory drugs. The animal recovered uneventfully. CONCLUSION: Intraocular foreign bodies secondary to gunshot wound should always be considered as a surgical emergency. Enucleation should be performed in cases having poor prognosis to avoid further complications especially in wild animals like Rhesus macaque.