Effects of dextrose prolotherapy on contusion-induced muscle injuries in mice

Abstract

Current treatment options for muscle injuries remain suboptimal and often result in delayed/incomplete recovery of damaged muscles. In this study, the effects of dextrose prolotherapy on inflammation and regeneration of skeletal muscles after a contusion injury were investigated. Mice were separated into five groups, including a normal control (NC), post-injury with no treatment (mass-drop injury, MDI), post-injury with 10% dextrose (MDI + 10% dextrose), post-injury with 20% dextrose (MDI + 20% dextrose), and post-injury with 30% dextrose (MDI + 30% dextrose). The gastrocnemius muscles of the mice were subjected to an MDI, and muscle samples were collected at 7 days post-injury. Results showed the serum creatine kinase (CK), blood urea nitrogen (BUN), creatinine (CREA), and low-density lipoprotein (LDH) of the MDI-alone group were significantly higher than those of the normal control group (p<0.05). However, levels of serum CK, BUN, CREA, and lactate dehydrogenase (LDH) significantly decreased with different concentrations of dextrose. In addition, dextrose suppressed the macrophage response (F4/80 protein decreased) and promoted muscle satellite cell regeneration (desmin protein increased). In conclusion, dextrose prolotherapy can effectively help repair muscles; therefore, it may be one of the methods for clinically treating muscle injuries.

Keywords: contusion; dextrose; diclofenac (DCF).; mass-drop injury (MDI); prolotherapy.

Conflict of interest statement

Competing Interests: The authors have declared that no competing interest exists.

Figures

Figure 1

(A) The schedule of dextrose prolotherapy treatment for contusion-induced muscle injuries in mice. (B) The gastrocnemius muscle of mice was subjected to a mass-drop injury (MDI). Mice were randomly assigned to six groups (8 mice/group), namely: (1) normal control (NC), animals injected with saline without injury; (2) MDI, animals injected with saline at 2 h post-injury; (3) MDI + 10% dextrose, animals injected with 10% dextrose at 2 h post-injury; (4) MDI + 20% dextrose, animals injected with 20% dextrose at 2 h post-injury; (5) MDI + 30% dextrose, animals injected with 30% dextrose at 2 h post-injury; and (6) MDI + DCF (diclofenac, an NSAID), animals administrated DCF at 2 h post-injury.

Figure 2

Effect of dextrose prolotherapy on serum blood urea nitrogen (BUN) on contusion-induced muscle injuries in mice. Mice were randomly assigned to five groups (8 mice/group), namely: (1) normal control (NC), animals injected with saline without injury; (2) mass-drop injury (MDI), animals injected with saline at 2 h post-injury; (3) MDI + 10% dextrose, animals injected with 10% dextrose at 2 h post-injury; (4) MDI + 20% dextrose, animals injected with 20% dextrose at 2 h post-injury; and (5) MDI + 30% dextrose, animals injected with 30% dextrose at 2 h post-injury. Data are the mean±SEM (n = 8 mice/group). Different letters indicate a significant difference at p<0.05 by a one-way ANOVA.

Figure 3

Effect of dextrose prolotherapy on serum creatinine (CREA) on contusion-induced muscle injuries in mice. Mice were randomly assigned to five groups (8 mice/group), namely: (1) normal control (NC), animals injected with saline without injury; (2) mass-drop injury (MDI), animals injected with saline at 2 h post-injury; (3) MDI + 10% dextrose, animals injected with 10% dextrose at 2 h post-injury; (4) MDI + 20% dextrose, animals injected with 20% dextrose at 2 h post-injury; and (5) MDI + 30% dextrose, animals injected with 30% dextrose at 2 h post-injury. Data are the mean±SEM (n = 8 mice/group). Different letters indicate a significant difference at p<0.05 by a one-way ANOVA.

Figure 4

Effect of dextrose prolotherapy on serum lactate dehydrogenase (LDH) on contusion-induced muscle injuries in mice. Mice were randomly assigned to five groups (8 mice/group), namely: (1) normal control (NC), animals injected with saline without injury; (2) mass-drop injury (MDI), animals injected with saline at 2 h post-injury; (3) MDI + 10% dextrose, animals injected with 10% dextrose at 2 h post-injury; (4) MDI + 20% dextrose, animals injected with 20% dextrose at 2 h post-injury; and (5) MDI + 30% dextrose, animals injected with 30% dextrose at 2 h post-injury. Data are the mean±SEM (n = 8 mice/group). Different letters indicate a significant difference at p<0.05 by a one-way ANOVA.

Figure 5

Effect of dextrose prolotherapy on serum creatine kinase (CK) on contusion-induced muscle injuries in mice. Mice were randomly assigned to five groups (8 mice/group), namely: (1) normal control (NC), animals injected with saline without injury; (2) mass-drop injury (MDI), animals injected with saline at 2 h post-injury; (3) MDI + 10% dextrose, animals injected with 10% dextrose at 2 h post-injury; (4) MDI + 20% dextrose, animals injected with 20% dextrose at 2 h post-injury; and (5) MDI + 30% dextrose, animals injected with 30% dextrose at 2 h post-injury. Data are the mean±SEM (n = 8 mice/group). Different letters indicate a significant difference at p<0.05 by a one-way ANOVA.

Figure 6

Effects of dextrose prolotherapy on pathological histology of (A) the liver, (B) muscles, (C) lungs, (D) kidneys, (E) heart, and (F) epididymal fat pad tissues on contusion-induced muscle injuries in mice. Mice were randomly assigned to seven groups (8 mice/group), namely: (1) vehicle + 30% dextrose, animals injected with 30% dextrose without injury; (2) vehicle control, animals injected with saline without injury; (3) mass-drop injury (MDI), animals injected with saline at 2 h post-injury; (4) MDI + 10% dextrose, animals injected with 10% dextrose at 2 h post-injury; (5) MDI + 20% dextrose, animals injected with 20% dextrose at 2 h post-injury; (6) MDI + 30% dextrose, animals injected with 30% dextrose at 2 h post-injury; and (7) MDI + DCF (diclofenac, an NSAID), animals administrated DCF at 2 h post-injury.

Figure 7

IHC staining of (A) F4/80 protein and (B) desmin protein in muscle tissues after contusion-induced muscle injuries in mice. Mice were randomly assigned to seven groups (8 mice/group), namely: (1) vehicle + 30% dextrose, animals injected with 30% dextrose without injury; (2) vehicle control, animals injected with saline without injury; (3) mass-drop injury (MDI), animals injected with saline at 2 h post-injury; (4) MDI + 10% dextrose, animals injected with 10% dextrose at 2 h post-injury; (5) MDI + 20% dextrose, animals injected with 20% dextrose at 2 h post-injury; (6) MDI + 30% dextrose, animals injected with 30% dextrose at 2 h post-injury; and (7) MDI + DCF (diclofenac, an NSAID), animals administrated DCF at 2 h post-injury.