Photobiomodulation therapy using a class 3b laser does not cause premature rupture of multivesicular liposomal bupivacaine suspensions in a cadaver model.

OBJECTIVE: To determine if photobiomodulation causes a premature release of liposomal bupivacaine (LB) suspensions. ANIMALS: A 25-kg mixed breed dog cadaver euthanized for reasons unrelated to this study. METHODS: In September 2022, a proximomedial tibial incision was made in a dog cadaver, and a tibial plateau leveling osteotomy plate was implanted. A stab incision was made one-half inch distal to the incision, and a tunnel to the plate was created prior to closure of the primary wound. A 3-cc anal sac catheter was advanced through the distal incision until the bulb rested against the face of the plate. Seven treatment groups of treatment power (watts) and total energy (joules/cm2) were defined as: A, 0.5 W, 2.0 J/cm2; B, 0.5 W, 4.0 J/cm2; C, 0.5 W, 6.0 J/cm2; D, 1.0 W, 2.0 J/cm2; E, 1.0 W, 4.0 J/cm2; F, 1.0 W, 6.0 J/cm2; and sham, 0.0 W, 0.0 J/cm2. Ten samples per group of 2 mL of LB were infused into a new catheter and treated percutaneously with a class 3b laser. All samples remained in the catheter for 12 seconds to reflect the longest treatment time. Post-treatment free bupivacaine concentrations were identified with high-performance liquid chromatography. RESULTS: The median free bupivacaine concentration was reported as: sham, 1.89 mg/mL; A, 1.93 mg/mL; B, 2.01 mg/mL; C, 2.05 mg/mL; D, 1.92 mg/mL; E, 2.03 mg/mL; and F, 2.00 mg/mL. There were no differences in median free bupivacaine concentrations between groups (P = .988). CLINICAL RELEVANCE: Concurrent LB and photobiomodulation are recommended during the postoperative period. The results of this proof-of-concept study suggest that concurrent use of LB and photobiomodulation may be safe, but in vivo studies at similar and stronger photobiomodulation settings are warranted.

The Drosophila drop-dead gene is required for eggshell integrity.

The eggshell of the fruit fly Drosophila melanogaster is a useful model for understanding the synthesis of a complex extracellular matrix. The eggshell is synthesized during mid-to-late oogenesis by the somatic follicle cells that surround the developing oocyte. We previously reported that female flies mutant for the gene drop-dead (drd) are sterile, but the underlying cause of the sterility remained unknown. In this study, we examined the role of drd in eggshell synthesis. We show that eggs laid by drd mutant females are fertilized but arrest early in embryogenesis, and that the innermost layer of the eggshell, the vitelline membrane, is abnormally permeable to dye in these eggs. In addition, the major vitelline membrane proteins fail to become crosslinked by nonreducible bonds, a process that normally occurs during egg activation following ovulation, as evidenced by their solubility and detection by Western blot in laid eggs. In contrast, the Cp36 protein, which is found in the outer chorion layers of the eggshell, becomes crosslinked normally. To link the drd expression pattern with these phenotypes, we show that drd is expressed in the ovarian follicle cells beginning in mid-oogenesis, and, importantly, that all drd mutant eggshell phenotypes could be recapitulated by selective knockdown of drd expression in the follicle cells. To determine whether drd expression was required for the crosslinking itself, we performed in vitro activation and crosslinking experiments. The vitelline membranes of control egg chambers could become crosslinked either by incubation in hyperosmotic medium, which activates the egg chambers, or by exogenous peroxidase and hydrogen peroxide. In contrast, neither treatment resulted in the crosslinking of the vitelline membrane in drd mutant egg chambers. These results indicate that drd expression in the follicle cells is necessary for vitelline membrane proteins to serve as substrates for peroxidase-mediated cross-linking at the end of oogenesis.

The Drosophila drop-dead gene is required for eggshell integrity.

The eggshell of the fruit fly Drosophila melanogaster is a useful model for understanding the synthesis of a complex extracellular matrix. The eggshell is synthesized during mid-to-late oogenesis by the somatic follicle cells that surround the developing oocyte. We previously reported that female flies mutant for the gene drop-dead ( drd ) are sterile, but the underlying cause of the sterility remained unknown. In this study, we examined the role of drd in eggshell synthesis. We show that eggs laid by drd mutant females are fertilized but arrest early in embryogenesis, and that the innermost layer of the eggshell, the vitelline membrane, is abnormally permeable to dye in these eggs. In addition, the major vitelline membrane proteins fail to become crosslinked by nonreducible bonds, a process that normally occurs during egg activation following ovulation, as evidenced by their solubility and detection by Western blot in laid eggs. In contrast, the Cp36 protein, which is found in the outer chorion layers of the eggshell, becomes crosslinked normally. To link the drd expression pattern with these phenotypes, we show that drd is expressed in the ovarian follicle cells beginning in mid-oogenesis, and, importantly, that all drd mutant eggshell phenotypes could be recapitulated by selective knockdown of drd expression in the follicle cells. To determine whether drd expression was required for the crosslinking itself, we performed in vitro activation and crosslinking experiments. The vitelline membranes of control egg chambers could become crosslinked either by incubation in hyperosmotic medium, which activates the egg chambers, or by exogenous peroxidase and hydrogen peroxide. In contrast, neither treatment resulted in the crosslinking of the vitelline membrane in drd mutant egg chambers. These results indicate that drd expression in the follicle cells is necessary for vitelline membrane proteins to serve as substrates for peroxidase-mediated cross-linking at the end of oogenesis.