The aim of this paper was to evaluate the application potential of CS-HTCC/GP-0.1%Chx thermosensitive hydrogel which was synthesized using chitosan (CS), quaternized CS, and alpha,beta-glycerophosphate (alpha,beta-GP) loading with 0.1% chlorhexidine (Chx) (w/v) for periodontal treatment. An aqueous solution of CS-HTCC/GP-0.1%Chx was transformed into hydrogel at 6 min when the temperature was increased to 37 degrees C. The scan electron microscopy (SEM) image of the gel was a porous, loose and crosslinked network. In vitro, Chx released over 18 h from the CS-HTCC/GP thermosensitive hydrogel in artificial saliva pH 6.8. Release rate could be controlled through adjustment of alpha,beta-GP or Chx concentration. CS-HTCC/GP-0.1%Chx thermosensitive hydrogel exhibited excellent inhibitory activity against primary periodontal pathogens. CS-HTCC/GP-0.1%Chx thermosensitive hydrogel had no acute toxicity; the maximum tolerated dose in rats was 400 mg/ml. All results indicated that CS-HTCC/GP-0.1%Chx thermosensitive hydrogel is a strong candidate as a local drug delivery system for periodontal treatment.