RESUMEN
BACKGROUND: It is believed that phosphocitrate (PC) exerts its disease-modifying effects on osteoarthritis (OA) by inhibiting the formation of crystals. However, recent findings suggest that PC exerts its disease-modifying effect, at least in part, through a crystal-independent action. This study sought to examine the disease-modifying effects of PC and its analogue PC-ß-ethyl ester (PC-E) on partial meniscectomy-induced OA and the structure-activity relationship. METHODS: Calcification- and proliferation-inhibitory activities were examined in OA fibroblast-like synoviocytes (FLSs) culture. Disease-modifying effects were examined using Hartley guinea pigs undergoing partial meniscectomy. Cartilage degeneration was examined with Indian ink, safranin-O, and picrosirius red. Levels of matrix metalloproteinase-13 (MMP-13), ADAM metallopeptidase with thrombospondin type 1 motif 5 (ADAMTS5), chemokine (C-C motif) ligand 5 (CCL5), and cyclooxygenase-2 (Cox-2) were examined with immunostaining. The effects of PC-E and PC on gene expressions in OA FLSs were examined with microarray. Results are expressed as mean ± standard deviation and analyzed using Student's t test or Wilcoxon rank sum test. RESULTS: PC-E was slightly less powerful than PC as a calcification inhibitor but as powerful as PC in the inhibition of OA FLSs proliferation. PC significantly inhibited cartilage degeneration in the partial meniscectomied right knee. PC-E was less powerful than PC as a disease-modifying drug, especially in the inhibition of cartilage degeneration in the non-operated left knee. PC significantly reduced the levels of ADAMTS5, MMP-13 and CCL5, whereas PC-E reduced the levels of ADAMTS5 and CCL5. Microarray analyses revealed that PC-E failed to downregulate the expression of many PC-downregulated genes classified in angiogenesis and inflammatory response. CONCLUSIONS: PC is a disease-modifying drug for posttraumatic OA therapy. PC exerts its disease-modifying effect through two independent actions: inhibiting pathological calcification and modulating the expression of many genes implicated in OA. The ß-carboxyl group of PC plays an important role in the inhibition of cartilage degeneration, little role in the inhibition of FLSs proliferation, and a moderate role in the inhibition of FLSs-mediated calcification.
Asunto(s)
Antirreumáticos/farmacología , Cartílago Articular/efectos de los fármacos , Citratos/farmacología , Meniscos Tibiales/cirugía , Osteoartritis/tratamiento farmacológico , Membrana Sinovial/efectos de los fármacos , Proteínas ADAM/genética , Proteínas ADAM/metabolismo , Animales , Antirreumáticos/química , Calcinosis/prevención & control , Cartílago Articular/metabolismo , Cartílago Articular/patología , Proliferación Celular/efectos de los fármacos , Células Cultivadas , Quimiocina CCL5/genética , Quimiocina CCL5/metabolismo , Citratos/química , Ciclooxigenasa 2/genética , Ciclooxigenasa 2/metabolismo , Modelos Animales de Enfermedad , Relación Dosis-Respuesta a Droga , Fibroblastos/efectos de los fármacos , Fibroblastos/metabolismo , Fibroblastos/patología , Regulación de la Expresión Génica , Cobayas , Masculino , Metaloproteinasa 13 de la Matriz/genética , Metaloproteinasa 13 de la Matriz/metabolismo , Estructura Molecular , Osteoartritis/etiología , Osteoartritis/genética , Osteoartritis/metabolismo , Osteoartritis/patología , Relación Estructura-Actividad , Membrana Sinovial/metabolismo , Membrana Sinovial/patologíaRESUMEN
The pre-, peri-, and postoperative care of animal surgical models used for testing tissue engineering/regenerative medicine product candidates includes the thoughtful consideration of several important factors. It must ensure the health and comfort of the animals and the success and reproducibility of the model. In order to reduce the number of animals needed in creating the model and to reduce costs, a preliminary evaluation of surgical procedures and instruments should be performed on cadavers. Once a minimal level of proficiency has been acquired, non-survival surgeries should be executed successfully before attempting survival surgeries. Planning ahead is crucial and will involve all aspects of the animal's care such as allowing the animal to become accustomed to soft foods (as in the case of gastrointestinal surgeries), planning appropriate pain management, and the use of positive reinforcement. We present specific examples of pre-, peri- and post-operative care of rodents using our experiences in developing tissue engineering products for kidney, esophagus, small intestine and lung.
Asunto(s)
Crianza de Animales Domésticos/métodos , Bienestar del Animal , Animales de Laboratorio , Medicina Regenerativa/métodos , Ingeniería de Tejidos/métodos , Animales , Atención Perioperativa/veterinaria , Cuidados Posoperatorios/veterinaria , Cuidados Preoperatorios/veterinaria , RatasRESUMEN
Regenerative constructs composed of synthetically sourced, biodegradable biomaterials seeded with smooth muscle-like cells have been leveraged to mediate regeneration of bladder and bladder-like neo-organs. Here, we describe how such constructs may be applied to catalyze regeneration of esophagus and small intestine in preclinical rodent models.
Asunto(s)
Esófago/citología , Intestino Delgado/citología , Medicina Regenerativa/métodos , Síndrome del Intestino Corto/terapia , Ingeniería de Tejidos/métodos , Animales , Esófago/lesiones , RatasRESUMEN
Recent successes in regenerative medicine and tissue engineering of bladder and bladder-like neo-organs have leveraged regenerative constructs composed of a biodegradable scaffold seeded with a population of smooth muscle cells. We have shown that such smooth muscle cells are isolatable from adipose and other sources alternate to the primary organ. We hypothesize that this regenerative platform is not limited to regeneration of bladder and bladder-like neo-organs, but rather represents a foundational technology platform broadly applicable for regeneration of laminarly organized hollow organs. Using esophagus as an illustrative example in support of this hypothesis, we demonstrate that patch constructs composed of adipose-derived smooth muscle cells seeded on a biodegradable matrix catalyze complete regeneration of the esophageal wall in a rodent model of esophageal injury. By implication, such regenerative constructs may potentially be used to mediate the regeneration of any laminarly organized tubular organ.
Asunto(s)
Esófago/fisiología , Regeneración/fisiología , Ingeniería de Tejidos/métodos , Andamios del Tejido , Vejiga Urinaria/fisiología , Implantes Absorbibles , Animales , Femenino , Miocitos del Músculo Liso/patología , Ratas , Ratas Endogámicas Lew , Medicina Regenerativa , Ingeniería de Tejidos/instrumentaciónRESUMEN
AIMS: To apply an organ regeneration platform technology of autologous smooth muscle cell/biomaterial combination products, previously demonstrated to be successful for urinary tissue regeneration, to the regeneration of the small intestine. MATERIALS & METHODS: Patch and tubular constructs were implanted in rodent small intestines and histologically evaluated over a time course for evidence of regeneration of the laminarly organized neo-mucosa and muscle layers. RESULTS: Laminarly organized neo-mucosa and muscle layer bundles are demonstrated as early as 8 weeks postimplantation. CONCLUSION: An organ regeneration technology platform of autologous smooth muscle cell/biomaterial combination products can be extended to the regeneration of the small intestine.