Early inflammatory response of knee ligaments to prolotherapy in a rat model

J Orthop Res. 2008 Jun;26(6):816-23. doi: 10.1002/jor.20600.


Prolotherapy is an alternative injection-based therapy for chronic musculoskeletal pain. Three different proliferants, D-glucose (dextrose), phenol-glucose-glycerine (P2G), and sodium morrhuate, used in prolotherapy are hypothesized to strengthen and reorganize chronically injured soft tissue and decrease pain through modulation of the inflammatory process. Our hypothesis is that commonly used prolotherapy solutions will induce inflammation (leukocyte and macrophage infiltration) in medial collateral ligaments (MCLs) compared to needlestick, saline injection, and no-injection controls. MCLs of 84 Sprague- Dawley rats were injected one time at both the tibial and femoral insertions. Immunohistochemistry (IHC) was used to determine the inflammatory response at three locations (tibial and femoral insertions and midsubstance) 6, 24, and 72 h after dextrose injection compared to saline- and no-injection controls and collagenase (positive control) (n = 4). qPCR was used to analyze gene expression 24 h postinjection (n = 4). Sodium morrhuate, P2G, and needlestick control were also investigated after 24 h (n = 4). In general, inflammation (CD43+, ED1+, and ED2+ cells) increased after prolotherapy injection compared to no-injection control but did not increase consistently compared to saline and needlestick control injections. This response varied by both location and proliferant. Inflammation was observed at 6 and 24 h postinjection but was resolved by 72 h compared to no-injection controls (p < 0.05). CD43+ leukocytes and ED2+ macrophages increased compared to needlestick and saline-injection control, respectively, 24 h postinjection (p < 0.05). Prolotherapy injections created an inflammatory response, but this response was variable and overall, not uniformly different from that caused by saline injections or needlestick procedures.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Arthritis / chemically induced*
  • Arthritis / pathology*
  • Biomarkers / metabolism
  • Disease Models, Animal
  • Gene Expression / immunology
  • Glucose / pharmacology
  • Glycerol / pharmacology
  • Leukosialin / metabolism
  • Macrophages / metabolism
  • Macrophages / pathology
  • Medial Collateral Ligament, Knee / immunology*
  • Medial Collateral Ligament, Knee / pathology*
  • Needlestick Injuries
  • Neutrophils / metabolism
  • Neutrophils / pathology
  • Phenol / pharmacology
  • Polymerase Chain Reaction
  • Rats
  • Rats, Sprague-Dawley
  • Sclerosing Solutions / pharmacology*
  • Sodium Chloride / pharmacology
  • Sodium Morrhuate / pharmacology


  • Biomarkers
  • Leukosialin
  • Sclerosing Solutions
  • Phenol
  • Sodium Chloride
  • Sodium Morrhuate
  • Glucose
  • Glycerol