Hydrogels are hydrophilic polymers with three-dimensional networks that can absorb
water without dissolving and good properties such as high flexibility, hydrophilicity and water
absorption. Hydrogels are possible to develop in medical application. Deproteinzed natural rubber
latex has been attracted to be manufactured for various medical devices to prevent allergenicity
caused by natural rubber allergen proteins. Protease enzyme has been used for deproteinzed
natural rubber latex production because it is able to degrade allergic proteins. This research was
prepared hydrogels from deproteinized natural rubber latex with gelatinized cassava starch, sinlek
starch and rice flour using sulphur and N, N’ -methylene bisacrylamide (MBA) as crosslinking
agent. It was found that hydrogel from deproteinized natural rubber latex with gelatinized rice
flour gave the best cast thin film. Then preparation hydrogel from deproteinized natural rubber
latex with gelatinized rice flour and Caesulpinia sappan Linn extracts crosslink with MBA
showed 114.50 % swelling and 59.10 % water content. Studied of microorganism counting, it was
found that hydrogel sample had not both of fungus and bacteria and protein measurement showed
70.41 mg/g protein of hydrogel with protease less than 73.48 mg/g protein of hydrogel without
protease in mixed extraction (1 % SDS with tris-HCl pH 6.8). The extraction of phosphate buffer
saline pH 7.4 in with and without protease showed 5.74 and 6.14 mg/g protein, respectively. Thus,
hydrogel from deproteinized natural rubber latex with gelatinized rice flour and Caesulpinia
sappan Linn extracts crosslinked with MBA has swelling percentage, good water content and
efficiency of bacterial inhibition to develop for wound dressing in medical application.